Cell Transplantation 26(1) Abstracts

Return to Cell Transplantation>

Cell Transplantation, Vol. 26, pp. 1-9, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
693437
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Identification of Donor Origin and Condition of Transplanted Islets In Situ in the Liver of a Type 1 Diabetic Recipient

Cornelis R. van der Torren,*1 Jessica S. Suwandi,*1 DaHae Lee,†‡1 Ernst-Jan T. van’t Wout,*1 Gaby Duinkerken,*1 Godelieve Swings,*1 Arend Mulder,* Frans H. J. Claas,* Zhidong Ling,‡1 Pieter Gillard,†‡1 Bart Keymeulen,‡1 Peterin’t Veld,‡1 and Bart O. Roep1

*Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands
†Department of Endocrinology, Universitair Ziekenhuis Gasthuisberg, Catholic University of Leuven, Leuven, Belgium
‡Diabetes Research Center, Brussels Free University-VUB, Brussels, Belgium
§Department of Diabetes Immunology, Beckman Research Institute at the City of Hope, Duarte, CA, USA

Transplantation of islet allografts into type 1 diabetic recipients usually requires multiple pancreas donors to achieve insulin independence. This adds to the challenges of immunological monitoring of islet transplantation currently relying on surrogate immune markers in peripheral blood. We investigated donor origin and infiltration of islets transplanted in the liver of a T1D patient who died of hemorrhagic stroke 4 months after successful transplantation with two intraportal islet grafts combining six donors. Immunohistological staining for donor HLA using a unique panel of human monoclonal HLA-specific alloantibodies was performed on liver cryosections after validation on cryopreserved kidney, liver, and pancreas and compared with auto- and alloreactive T-cell immunity in peripheral blood. HLA-specific staining intensity and signal-to-noise ratio varied between tissues from very strong on kidney glomeruli, less in liver, kidney tubuli, and endocrine pancreas to least in exocrine pancreas, complicating the staining of inflamed islets in an HLA-disparate liver. Nonetheless, five islets from different liver lobes could be attributed to donors 1, 2, and 5 by staining patterns with multiple HLA types. All islets showed infiltration with CD8+ cytotoxic T cells that was mirrored by progressive alloreactive responses in peripheral blood mononuclear cells (PBMCs) to donors 1, 2, and 5 after transplantation. Stably low rates of peripheral islet autoreactive T-cell responses after islet infusion fit with a complete HLA mismatch between grafts and recipient and exclude the possibility that the islet-infiltrating CD8 T cells were autoreactive. HLA-specific immunohistochemistry can identify donor origin in situ and differentiate graft dysfunction and immunological destruction.

Key words: Type 1 diabetes; Islet transplantation; Autoimmunity; Alloreactivity

Received October 10, 2016; final acceptance October 10, 2016. Online prepub date: October 10, 2016.
1JDRF Center for Beta Cell Therapy in Diabetes.
Address correspondence to Professor Bart O. Roep, Ph.D., Department of Immunohaematology and Blood Transfusion, E3-Q, Leiden University Medical Center, P.O. Box 9600, NL-2300 RC Leiden, The Netherlands. Tel: +31-71-526 3800; Fax: +31-71-526 5267; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 11-21, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692096
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Physiologic Doses of Bilirubin Contribute to Tolerance of Islet Transplants by Suppressing the Innate Immune Response

Christopher A. Adin,* Zachary C. VanGundy,† Tracey L. Papenfuss,† Feng Xu,† Mostafa Ghanem,† Jonathan Lakey,‡ and Gregg A. Hadley§

*Department of Veterinary Clinical Sciences, North Carolina State University, Raleigh, NC, USA
†Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
‡Department of Surgery, University of California, Irvine, Irvine, CA, USA
§Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH, USA

Bilirubin has been recognized as a powerful cytoprotectant when used at physiologic doses and was recently shown to have immunomodulatory effects in islet allograft transplantation, conveying donor-specific tolerance in a murine model. We hypothesized that bilirubin, an antioxidant, acts to suppress the innate immune response to islet allografts through two mechanisms: 1) by suppressing graft release of damage-associated molecular patterns (DAMPs) and inflammatory cytokines, and 2) by producing a tolerogenic phenotype in antigen-presenting cells. Bilirubin was administered intraperitoneally before pancreatic procurement or was added to culture media after islet isolation in AJ mice. Islets were exposed to transplant-associated nutrient deprivation and hypoxia. Bilirubin significantly decreased islet cell death after isolation and hypoxic stress. Bilirubin supplementation of islet media also decreased the release of DAMPs (HMGB1), inflammatory cytokines (IL-1
β and IL-6), and chemokines (MCP-1). Cytoprotection was mediated by the antioxidant effects of bilirubin. Treatment of macrophages with bilirubin induced a regulatory phenotype, with increased expression of PD-L1. Coculture of these macrophages with splenocytes led to expansion of Foxp3+ Tregs. In conclusion, exogenous bilirubin supplementation showed cytoprotective and antioxidant effects in a relevant model of islet isolation and hypoxic stress. Suppression of DAMP release, alterations in cytokine profiles, and tolerogenic effects on macrophages suggest that the use of this natural antioxidant may provide a method of preconditioning to improve outcomes after allograft transplantation.

Key words: Islet; Damage-associated molecular patterns (DAMPs); HMGB1; Bilirubin; Innate immune response

Received April 7, 2016; final acceptance September 23, 2016. Online prepub date: July 7, 2016.
Address correspondence to Associate Professor Christopher A. Adin, D.V.M., D.A.C.V.S., College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27606, USA. Tel: (919) 513-6050; Fax: (919) 513-6715; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 23-32, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
693022
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Active Subjects With Autoimmune Type 1 Diabetes Have Better Metabolic Profiles Than Sedentary Controls

M. Adamo,*† R. Codella,*†‡ F. Casiraghi,† A. Ferrulli,† C. Macrì,† E. Bazzigaluppi,§ I. Terruzzi,¶ L. Inverardi,‡ C. Ricordi,‡ and L. Luzi*†‡

*Department of Biomedical Sciences for Health, Universita degli Studi di Milano, Milan, Italy
†Metabolism Research Center, IRCCS Policlinico San Donato, San Donato Milanese, Italy
‡Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA
§Laboraf-Diagnostic Research, San Raffaele Scientific Institute, Milan, Italy
¶Division of Metabolic and Cardiovascular Science, Metabolism, Nutrigenomics and Cellular Differentiation Unit, San Raffaele Scientific Institute, Milan, Italy

Previous studies in humans with type 1 diabetes mellitus (T1D) and in nonobese diabetic mice have investigated the beneficial immunomodulatory potential of aerobic physical activity. Performing high volume of aerobic exercise may favorably regulate autoimmunity in diabetes. We tested whether increased physical activity is a self-sufficient positive factor in T1D subjects. During a 3-month observational period, active (six males; 40.5
± 6.1 years; BMI: 24.5 ± 2.1) and sedentary (four males, three females; 35.9 ± 8.9 years; BMI: 25.7 ± 3.8) T1D individuals on insulin pump therapy were studied for metabolic, inflammatory, and autoimmune parameters. At baseline and at the end of a 3-month period, glycosylated hemoglobin (HbA1c), autoantibodies (anti-GAD, anti-ZnT8, anti-IA2, and ICA) and proinflammatory cytokines (IL-6 and TNF-α) were evaluated. During the third month of the period, physically active T1D patients showed a significant reduction in the average glucose levels (−9%, p = 0.025, by CGM) compared to the first month values, and even their hyperglycemic episodes (>180 mg/dl) diminished significantly (−24.2%, p = 0.032 vs. first month). Moreover, active T1D subjects exhibited an improved body composition with respect to sedentary controls. No significant changes were detected as to the autoimmune and inflammatory profiles. This study confirms the beneficial role of physical exercise associated with insulin pump therapy in order to improve metabolic control in individuals with T1D. These preliminary positive observations need to be challenged in a prolonged interventional follow-up.

Key words: Physical activity; Type 1 diabetes (T1D); Continuous glucose monitoring (CGM); Insulin pump therapy

Received July 5, 2016; final acceptance October 19, 2016. Online prepub date: September 22, 2016.
Address correspondence to Roberto Codella, Ph.D., Department of Biomedical Sciences for Health, Universita degli Studi di Milano, Via F.lli Cervi 93, 20090 Segrate (MI), Italy. Tel: +39-02-503-30300; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 33-44, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692834
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Effect of Manufacturing Procedures on Human Islet Isolation From Donor Pancreata Standardized by the North American Islet Donor Score

Chun-Chieh Yeh,*† Ling-jia Wang,* James J. McGarrigle,* Yong Wang,* Chien-Chang Liao,‡ Mustafa Omami,* Arshad Khan,* Mohammad Nourmohammadzadeh,* Joshua Mendoza-Elias,* Benjamin McCracken,* Enza Marchese,* Barbara Barbaro,* and Jose Oberholzer*

*Islet Transplantation Program, Division of Transplantation, Department of Surgery, University of Illinois at Chicago, Chicago, IL, USA
†Department of Surgery, China Medical University Hospital, China Medical University, Taichung, Taiwan
‡Department of Anesthesiology and Health Policy Research Center, Taipei Medical University Hospital, Taipei, Taiwan

This study investigates manufacturing procedures that affect islet isolation outcomes from donor pancreata standardized by the North American Islet Donor Score (NAIDS). Islet isolations performed at the University of Illinois, Chicago, from pancreata with NAIDS
³65 were investigated. The research cohort was categorized into two groups based on a postpurification yield either greater than (group A) or less than (group B) 400,000 IEQ. Associations between manufacturing procedures and islet isolation outcomes were analyzed using multivariate logistic or linear regressions. A total of 119 cases were retrieved from 630 islet isolations performed since 2003. Group A is composed of 40 cases with an average postpurified yield of 570,098 IEQ, whereas group B comprised 79 cases with an average yield of 235,987 IEQ. One third of 119 cases were considered successful islet isolations that yielded >400,000 IEQ. The prepurifiedand postpurified islet product outcome parameters were detailed for future reference. The NAIDS (>80 vs. 65–80) [odds ratio (OR): 2.91, 95% confidence interval (CI): 1.27–6.70], cold ischemic time (£10 vs. >10 h) (OR: 3.68, 95% CI: 1.61–8.39), and enzyme perfusion method (mechanical vs. manual) (OR: 2.38, 95% CI: 1.01–5.56) were independent determinants for postpurified islet yield ³400,000 IEQ. The NAIDS (>80, p < 0.001), cold ischemic time (£10 h, p < 0.05), increased unit of collagenase (p < 0.01), and pancreatic duct cannulation time (<30 min, p < 0.01) all independently correlated with better islet quantity parameters. Furthermore, cold ischemic time (£10 h, p < 0.05), liberase MTF (p < 0.001), increased unit of collagenase (p < 0.05), duct cannulation time (<30 min, p < 0.05), and mechanical enzyme perfusion (p < 0.05) were independently associated with better islet morphology score. Analysis of islet manufacturing procedures from the pancreata with standardized quality is essential in identifying technical issues within islet isolation. Adequate processing duration in each step of islet isolation, using liberase MTF, and mechanical enzyme perfusion all affect isolation outcomes.

Key words: Islet of Langerhans; Human islet isolation; North American Islet Donor Score (NAIDS); Good Manufacture Practice; Outcome parameters

Received May 5, 2016; final acceptance October 6, 2016. Online prepub date: August 12, 2016.
Address correspondence to Ling-jia Wang, Islet Laboratory, Division of Abdominal Organ Transplantation, Department of Surgery, University of Chicago, Rm 029, 910 E 58th Street, Chicago, IL 60637, USA. Tel: +1-773-834-8143; Fax: +1-773-702-7195; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jose Oberholzer, Department of Surgery/Transplant, University of Illinois at Chicago, Room 502, 840 S Wood Street, Chicago, IL 60612, USA. Tel: 312-996-6771; Fax: 312-413-7961; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 45-61, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692726
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Transplantation of Human Placenta-Derived Mesenchymal Stem Cells Alleviates Critical Limb Ischemia in Diabetic Nude Rats

Lu Liang,*† Zongjin Li,* Tao Ma,*† Zhibo Han,‡ Wenjing Du,‡ Jie Geng,† Honghong Jia,† Meng Zhao,† Jimin Wang,† Bingjing Zhang,† Jie Feng,† Lanzhen Zhao,† Alain Rupin,§ Youwei Wang,* and Zhong Chao Han*†‡

*Beijing Institute of Stem Cells, Health & Biotech Co., Beijing, P.R. China
†National Engineering Research Center of Cell Products, Tianjin, P.R. China
‡State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, P.R. China
§Servier Phamaceutical Co., Ltd., Suresnes, France

Neovasculogenesis induced by stem cell therapy is an innovative approach to improve critical limb ischemia (CLI) in diabetes. Mesenchymal stem cells (MSCs) are ideal candidates due to their angiogenic and immunomodulatory features. The aim of this study is to determine the therapeutic effects of human placenta-derived MSCs (P-MSCs) on diabetic CLI, with or without exogenous insulin administration, and the underlying mechanism of any effect. A series of in vitro experiments were performed to assess the stemness and vasculogenic activity of P-MSCs. P-MSCs were intramuscularly injected at two different doses with and without the administration of insulin. The efficacy of P-MSC transplantation was evaluated by ischemia damage score, ambulatory score, laser Doppler perfusion image (LDPI), capillary, and vascular density. In vivo imaging was applied to track the implanted P-MSCs. In vivo differentiation and in situ secretion of angiogenic cytokines were determined. In vitro experimental outcomes showed the differentiation potential and potent paracrine effect of P-MSCs. P-MSCs survived in vivo for at least 3 weeks and led to the acceleration of ischemia recovery, due to newly formed capillaries, increased arterioles, and secretion of various proangiogenic factors. P-MSCs participate in angiogenesis and vascularization directly through differentiation and cytokine expression.

Key words: Placenta-derived mesenchymal stem cells (P-MSCs); Critical limb ischemia (CLI); Angiogenesis; Cell therapy

Received February 29, 2016; final acceptance October 14, 2016. Online prepub date: August 5, 2016.
Address correspondence to Zhong Chao Han, 288 Nan Jing Road, Heping District, Tianjin 300080, P.R. China. Tel: 13802030084; Fax: 86-022-66210734; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 63-70, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692825
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Excluding Anti-cytomegalovirus Immunoglobulin M-Positive Cord Blood Units Has a Minimal Impact on the Korean Public Cord Blood Bank Inventory

Sue Shin,*†‡1 Eun Youn Roh,*†‡1 Sohee Oh,§ Eun Young Song,* Eui Chong Kim,* and Jong Hyun Yoon*†‡

*Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, South Korea
†Department of Laboratory Medicine, Boramae Hospital, Seoul, South Korea
‡Seoul Metropolitan Government Public Cord Blood Bank (ALLCORD), Seoul, South Korea
§Department of Biostatistics, Boramae Hospital, Seoul, South Korea

Cord blood units (CBUs) for transplantation should be free of communicable disease and must contain a specific amount of total nucleated cells and CD34+ cells. Although posttransplantation cytomegalovirus (CMV) infections are from latent infection in patients, ensuring CMV-free CBUs by performing CMV-specific IgM and nucleic acid amplification testing (NAT) is one of the mandatory procedures for the safety of CBUs. However, the exclusion policies (based on these test results) vary among nations and institutions. We tested 28,000 processed CBUs between May 2006 and June 2014. The cord blood leukocytes from CMV IgM+ samples were then subjected to NAT. The total nucleated cell and CD34+ cell counts were measured for each CBU, and the results were compared to the CMV IgM and IgG results. The seroprevalence of CMV among pregnant women was 98.1% (18,459/18,818) for IgG and 1.7% (441/25,293) for IgM. The concentration and the total number of CD34+ cells were significantly higher in CBUs from IgMmothers compared to those from IgM+ mothers (72.4/μl vs. 57.2/μl, respectively,
p < 0.0001; 1.45 × 106/unit vs. 1.15 × 106/unit, respectively, p < 0.0001). Among CBUs with positive CMV IgM in their mothers’ plasma or cord blood plasma, only 0.58% of the samples (3:517) had a positive NAT. The number of excluded CBUs from inventory due to positive CMV IgM in the cord blood was 54 of 18,326 (0.3%). For inventory purposes, it is appropriate to remove CBUs with positive cord blood CMV IgM findings irrespective of the NAT status as well as positive maternal CMV IgM in South Korea.

Key words: Cytomegalovirus (CMV); Cord blood; Banking; Inventory

Received October 17, 2015; final acceptance September 20, 2016. Online prepub date: August 12, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Jong Hyun Yoon, M.D., Department of Laboratory Medicine, Seoul National University Boramae Hospital, 20 Boramaero-5-gil, Dongjak-gu, Seoul 156-707, South Korea. Tel: 82-2-870-2601; Fax: 82-2-870-2826; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 71-81, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692609
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Control of IBMIR Induced by Fresh and Cryopreserved Hepatocytes by Low Molecular Weight Dextran Sulfate Versus Heparin

Elisabet Gustafson,* Sana Asif,† Huda Kozarcanin,† Graciela Elgue,† Staffan Meurling,* Kristina N. Ekdahl,‡ and Bo Nilsson†

*Department of Women’s and Children’s Health, Division of Pediatric Surgery, Uppsala University Hospital, Uppsala, Sweden
†Department of Immunology, Genetics and Pathology (IGP), Uppsala University, Uppsala, Sweden
Linnæus Center of Biomaterials Chemistry, Linnæus University, Kalmar, Sweden

Rapid destruction of hepatocytes after hepatocyte transplantation has hampered the application of this procedure clinically. The instant blood-mediated inflammatory reaction (IBMIR) is a plausible underlying cause for this cell loss. The present study was designed to evaluate the capacity of low molecular weight dextran sulfate (LMW-DS) to control these initial reactions from the innate immune system. Fresh and cryopreserved hepatocytes were tested in an in vitro whole-blood model using ABO-compatible blood. The ability to elicit IBMIR and the capacity of LMW-DS (100 μg/ml) to attenuate the degree of activation of the cascade systems were monitored. The effect was also compared to conventional anticoagulant therapy using unfractionated heparin (1 IU/ml). Both fresh and freeze–thawed hepatocytes elicited IBMIR to the same extent. LMW-DS reduced the platelet loss and maintained the cell counts at the same degree as unfractionated heparin, but controlled the coagulation and complement systems significantly more efficiently than heparin. LMW-DS also attenuated the IBMIR elicited by freeze–thawed cells. Therefore, LMW-DS inhibits the cascade systems and maintains the cell counts in blood triggered by both fresh and cryopreserved hepatocytes in direct contact with ABO-matched blood. LMW-DS at a previously used and clinically applicable concentration (100 μg/ml) inhibits IBMIR in vitro and is therefore a potential IBMIR inhibitor in hepatocyte transplantation.

Key words: Innate immunity; IBMIR; Thromboinflammation; Hepatocyte transplantation; Low molecular weight dextran sulfate (LMW-DS)

Received May 22, 2016; final acceptance September 28, 2016. Online prepub date: July 22, 2016.
Address correspondence to Elisabet Gustafson, Division of Pediatric Surgery, Department of Women’s and Children’s Health, Uppsala University Children’s Hospital, SE-751 85 Uppsala, Sweden. Tel: +46186990330; Fax: +46186115905; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 83-93, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692221
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Poor Mobilization in T-Cell-Deficient Nude Mice Is Explained by Defective Activation of Granulocytes and Monocytes

Marcin Wysoczynski,*† Mateusz Adamiak,* Malwina Suszynska,* Ahmed Abdel-Latif,‡ Janina Ratajczak,* and Mariusz Z. Ratajczak

*Stem Cell Institute at James Graham Brown Cancer Center, University of Louisville, KY, USA
†Institute of Molecular Cardiology, University of Louisville, KY, USA
‡Division of Cardiovascular Medicine, Gill Heart Institute, University of Kentucky, Lexington, KY, USA
§Department of Regenerative Medicine, Warsaw Medical University, Warsaw, Poland

It has been reported that both SCID mice and SCID patients poorly mobilize hematopoietic stem/progenitor cells (HSPCs) in response to granulocyte colony-stimulating factor (G-CSF). This defect has been proposed to result from a lack of naturally occurring IgM immunoglobulins to trigger activation of the complement cascade (ComC) and release of C5 cleavage fragments crucial in the mobilization process. However, SCID individuals also have T-cell deficiency, and T cells have been shown to modulate trafficking of HSPCs. To learn more about the role of T lymphocytes, we performed mobilization studies in T-lymphocyte-deficient nude mice and found that these mice respond poorly to G-CSF and zymosan but are normal mobilizers in response to AMD3100. Since nude mice have normal levels of IgM immunoglobulins in peripheral blood and may activate the ComC, we focused on the potential involvement of Gr1+ granulocytes and monocytes, which show defective maturation in these animals. Using a nude mouse mobilization model, we found further support for the proposition that proper function of Gr1+ cells is crucial for optimal mobilization of HSPCs.

Key words: Complement; Granulocyte degranulation; Stem cell mobilization

Received March 15, 2016; final acceptance September 22, 2016. Online prepub date: July 18, 2016.
Address correspondence to Professor Mariusz Z. Ratajczak, M.D., Ph.D., Stem Cell Institute at James Graham Brown Cancer Center University of Louisville, 500 South Floyd Street, Louisville, KY 40202, USA. Tel: (502) 852-1788; Fax: (502) 852-3032; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 95-102, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692816
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Differences in Tfh Cell Response Between the Graft and Spleen With Chronic Allograft Nephropathy

Jian Shi,* Xianlin Xu,* Fengbao Luo,* Qianqian Shi,* Xiaozhou He,* and Ying Xia†

*The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu, P.R. China
†The University of Texas McGovern Medical School, Houston, TX, USA

The aim of this study was to investigate follicular helper T (Tfh) cell response and its difference between renal graft and spleen in a rat renal transplantation model undergoing chronic allograft nephropathy (CAN). Orthotopical kidney transplantations were performed on Fischer (F344) rats and transplanted to Lewis rats, using syngeneic Lewis–Lewis grafts as controls. Tissue samples were collected at 8 weeks posttransplantation. The status of Tfh cell response was assessed by measuring the levels of transcription factor B-cell lymphoma 6 (Bcl-6), interleukin-21 (IL-21), chemokine receptor type 5 (CXCR5), and B-cell-activating factor belonging to the TNF family (BAFF). Tfh cell response was upregulated in both renal graft and spleen of the CAN group compared to the control group. However, Tfh cell response of the spleen was weaker than that of the graft, which was possibly related to the upregulation of splenic Tregactivation. Also, the difference between two tissues was partially associated with the different expressions of tristetraprolin (TTP)/IL-10. Our data help improve our understanding of the role of Tfh cell response in the body with CAN and may provide a valuable clue for better treatment of CAN.

Key words: Tfh cell response; Chronic allograft nephropathy (CAN); Renal graft; Spleen

Received June 8, 2016; final acceptance October 5, 2016. Online prepub date: August 12, 2016.
Address correspondence to Xiaozhou He, Department of Urology, The Third Affiliated Hospital of Soochow University, Changzhou, 213000 Jiangsu Province, P.R. China. Tel: +86-0519-68870351; Fax: +86-0519-68870351; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Ying Xia, Department of Neurosurgery, The University of Texas McGovern Medical School, Houston, TX 7030, USA. Tel: +001-713-500-6288; Fax: +001-713-500-0601; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 103-113, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692249
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Improved Transplanted Stem Cell Survival in a Polymer Gel Supplemented With Tenascin C Accelerates Healing and Reduces Scarring of Murine Skin Wounds

Cecelia C. Yates,*†‡§1 Austin Nuschke,*‡1 Melanie Rodrigues,¶ Diana Whaley,*§ Jason J. Dechant,† Donald P. Taylor,‡#**†† and Alan Wells*‡§††

*Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
†Department of Health Promotion and Development, University of Pittsburgh School of Nursing, Pittsburgh, PA, USA
‡McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA
§Veterans Affairs Medical Center, Pittsburgh, PA, USA
¶Department of Plastic Surgery, Stanford University, CA, USA
#Department of Biomedical Informatics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
**Department of Plastic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
††Department of Bioengineering, University of Pittsburgh Swanson School of Engineering, Pittsburgh, PA, USA

Mesenchymal stem cells (MSCs) remain of great interest in regenerative medicine because of their ability to home to sites of injury, differentiate into a variety of relevant lineages, and modulate inflammation and angiogenesis through paracrine activity. Many studies have found that despite the promise of MSC therapy, cell survival upon implant is highly limited and greatly reduces the therapeutic utility of MSCs. The matrikine tenascin C, a protein expressed often at the edges of a healing wound, contains unique EGF-like repeats that are able to bind EGFR at low affinities and induce downstream prosurvival signaling without inducing receptor internalization. In this study, we utilized tenascin C in a collagen/GAG-based polymer (TPolymer) that has been shown to be beneficial for skin wound healing, incorporating human MSCs into the polymer prior to application to mouse punch biopsy wound beds. We found that the TPolymer was able to promote MSC survival for 21 days in vivo, leading to associated improvements in wound healing such as dermal maturation and collagen content. This was most marked in a model of hypertrophic scarring, in which the scar formation was limited. This approach also reduced the inflammatory response in the wound bed, limiting CD3e+ cell invasion by approximately 50% in the early wound-healing process, while increasing the numbers of endothelial cells during the first week of wound healing as well. Ultimately, this matrikine-based approach to improving MSC survival may be of great use across a variety of cell therapies utilizing matrices as delivery vehicles for cells.

Key words: Cellular therapy; PEG-polymer; Mesenchymal stem cells (MSCs); Extracellular matrix; Collagen; Tenascin C; Dermal wound healing

Received February 4, 2016; final acceptance October 25, 2016. Online prepub date: July 22, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Alan Wells, M.D., D.M.Sc., Department of Pathology, University of Pittsburgh, 3550 Terrace Street, Scaife Hall, S-713, Pittsburgh, PA 15261, USA. Tel: 412-647-7813; Fax: 412-647-8567; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Cecelia C. Yates, Ph.D., University of Pittsburgh School of Nursing, 3500 Victoria Street, Victoria Building, 458A, Pittsburgh, PA 15261, USA. Tel: 412-624-5728; Fax: 412-624-8521; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 115-123, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
693338
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells

Ah young Kim, Yongsun Kim, Seung Hoon Lee, Yongseok Yoon, Wan-Hee Kim, and Oh-Kyeong Kweon

BK21 PLUS Program for Creative Veterinary Science Research, Research Institute for Veterinary Science and College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea

Osteogenically differentiated cell sheet techniques using mesenchymal stem cells (MSCs) are available to stimulate bone regeneration. The advantage of the cell sheet technique is delivering live cells effectively into the focal region. We developed a novel osteogenic cell sheet technique by adding gelatin to osteogenic cell medium. Gelatin-induced osteogenic cell sheets (GCSs) were compared to conventional osteogenic cell sheets (OCSs). Undifferentiated MSCs (UCs) were used as a control. The morphology of these cell sheets was evaluated microscopically and histologically. The time-dependent cell proliferation rate was estimated by DNA quantification. The expression of osteogenic gene markers and the number of calcium depositions were assessed by quantitative real-time polymerase chain reaction and Alizarin red S (ARS) staining, respectively. GCSs were thicker and stronger than OCSs. GCSs showed a significantly higher cell proliferation rate compared to OCSs (
p < 0.05). GCSs exhibited significantly higher upregulation of BMP-7 mRNA compared to OCSs (p < 0.05). Both GCSs and OCSs showed negative ARS reactivity on day 10, but only GCSs showed positive ARS reactivity on day 21. With this technique, we observed active cell proliferation with abundant ECM and upregulation of osteogenic bone markers, and our results suggest that GCSs could be promising for therapeutic applications in bone regeneration.

Key words: Bone regeneration; Gelatin; Mesenchymal stem cells (MSCs); Osteogenic cell sheets (OCSs)

Received September 6, 2016; final acceptance October 21, 2016. Online prepub date: October 7, 2016.
Address correspondence to Oh-Kyeong Kweon, Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul 08826, Republic of Korea. Tel: +82-2-880-1248; Fax: +82-2-888-2866; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 125-133, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692717
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Biliary Polyunsaturated Fatty Acids and Telocytes in Gallstone Disease

Artur Pasternak,*† Jolanta Bugajska,‡ Mirosław Szura,§ Jerzy A. Walocha,† Andrzej Matyja,* Mariusz Gajda,¶ Krystyna Sztefko,‡ and Krzysztof Gil#

*First Department of General, Oncological and Gastrointestinal Surgery, Jagiellonian University Medical College, Krakow, Poland
†Department of Anatomy, Jagiellonian University Medical College, Krakow, Poland
‡Department of Clinical Biochemistry, Jagiellonian University Medical College, Krakow, Poland
§Department of Experimental and Clinical Surgery, Faculty of Health Sciences, Jagiellonian University Medical College, Krakow, Poland
¶Department of Histology, Jagiellonian University Medical College, Krakow, Poland
#Department of Pathophysiology, Jagiellonian University Medical College, Krakow, Poland

It has been reported that intake of
ω-3 polyunsaturated fatty acids (PUFAs) reduces the risk of coronary heart disease. It also influences bile composition, decreasing biliary cholesterol saturation in the bile of patients with gallstones. In addition to bile composition disturbances, gallbladder hypomotility must be a cofactor in the pathogenesis of cholelithiasis, as it leads to the prolonged nucleation phase. Our current knowledge about gallbladder motility has been enhanced by the study of a population of newly described interstitial (stromal) cells—telocytes (TCs). The purpose of this study was to determine whether TC loss, reported by our team recently, might be related to bile lithogenicity, expressed as cholesterol saturation index or the difference in biliary PUFA profiles in patients who suffer from cholecystolithiasis and those not affected by this disease. We determined biliary lipid composition including the fatty acid composition of the phospholipid species in bile. Thus, we investigated whether differences in biliary fatty acid profiles (ω-3 PUFA and ω-6 PUFA) in gallbladder bile may influence its lithogenicity and the quantity of TCs within the gallbladder wall. We conclude that the altered PUFA concentrations in the gallbladder bile, with elevation of ω-6 PUFA, constitute important factors influencing TC density in the gallbladder wall, being one of the possible pathophysiological components for the gallstone disease development. This study established that altered bile composition in patients with cholelithiasis may influence TC quantity within the gallbladder muscle, and we concluded that reduction in TC number may be a consequence of the supersaturated bile toxicity, while some other bile components (ω-3 PUFA, glycocholic, and taurocholic acids) may exert protective effects on TC and thus possibly influence the mechanisms regulating gallbladder and extrahepatic bile duct motility. Thus, ω-3 PUFA may represent a possible option to prevent formation of cholesterol gallstones.

Key words: Telocytes (TCs); Interstitial Cajal-like cells; Gallstones; Cholesterol saturation index (CSI); Bile lithogenicity; Polyunsaturated fatty acids (PUFAs); Biliary fatty acids

Received June 13, 2016; final acceptance September 26, 2016. Online prepub date: August 5, 2016.
Address correspondence to Artur Pasternak, M.D., Ph.D., Department of Anatomy, Jagiellonian University Medical College, Kopernika 12 Street, 31-034 Krakow, Poland. Tel: + 4812-4229511; Fax: + 4812-4229511; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 135-143, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692942
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Cellular Evidence of Telocytes as Novel Interstitial Cells Within the Magnum of Chicken Oviduct

Ping Yang,* Xudong Zhu,† Lingling Wang,* Nisar Ahmed,* Yufei Huang,* Hong Chen,* Qian Zhang,* Shakeeb Ullah,* Tengfei Liu,* Dawei Guo,* Sarfaraz Ahmed Brohi,* and Qiusheng Chen*

*College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, P.R. China
†College of Sciences, Nanjing Agricultural University, Nanjing, P.R. China

Telocytes are a novel type of interstitial cell that has been identified in many organs of mammals, but there is little information available on these cells in avian species. This study shows the latest findings associated with telocytes in the muscular layer and lamina propria of the magnum of chicken oviduct analyzed by transmission electron microscopy. Telocytes are characterized by telopodes, which are thin and long prolongations, and a small amount of cytoplasm rich with mitochondria. Spindle- or triangular-shaped telocytes were detected at various locations in the magnum. In the muscular layer, telocytes have direct connection with smooth muscle cells. The cell body of telocytes along with their long telopodes mainly exists in the interstitial space between the smooth muscle bundles, whereas large numbers of short telopodes are scattered in between the smooth muscle cells. In the lamina propria, extremely long telopodes are twisting around each other and are usually collagen embedded. Both in the lamina propria and muscular layer, telocytes have a close relationship with other cell types, such as immune cells and blood vessels. Telopodes appear with dichotomous branching alternating between the podom and podomer, forming a 3D network structure with complex homo- and heterocellular junctions. In addition, a distinctive size of the vesicles is visible around the telopodes and may be released from telopodes because of the close relation between the vesicle and telopode. All characteristics of telocytes in the magnum indicate that telocytes may play a potential, but important, role in the pathogenesis of oviduct diseases.

Key words: Telocytes (TCs); Oviduct; Chicken; Ultrastructure

Received July 13, 2016; final acceptance October 19, 2016. Online prepub date: September 1, 2016.
Address correspondence to Qiusheng Chen, College of Veterinary Medicine in Nanjing Agricultural University, Nanjing 210095, P.R. China. Tel: +86-25-8439530; Fax: +86-25-84398669; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 145-156, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692861
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Pivotal Role of Brain-Derived Neurotrophic Factor Secreted by Mesenchymal Stem Cells in Severe Intraventricular Hemorrhage in Newborn Rats

So Yoon Ahn,*†1 Yun Sil Chang,*†‡1 Dong Kyung Sung,*† Se In Sung,* Jee-Yin Ahn,§ and Won Soon Park*†‡

*Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea
†Stem Cell and Regenerative Medicine Institute, Samsung Medical Center, Seoul, South Korea
‡Department of Health Sciences and Technology, SAIHST, Sungkyunkwan University, Seoul, South Korea
§Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon, South Korea

Mesenchymal stem cell (MSC) transplantation protects against neonatal severe intraventricular hemorrhage (IVH)-induced brain injury by a paracrine rather than regenerative mechanism; however, the paracrine factors involved and their roles have not yet been delineated. This study aimed to identify the paracrine mediator(s) and to determine their role in mediating the therapeutic effects of MSCs in severe IVH. We first identified significant upregulation of brain-derived neurotrophic factor (BDNF) in MSCs compared with fibroblasts, in both DNA and antibody microarrays, after thrombin exposure. We then knocked down BDNF in MSCs by transfection with small interfering (si)RNA specific for human BDNF. The therapeutic effects of MSCs with or without BDNF knockdown were evaluated in vitro in rat neuronal cells challenged with thrombin, and in vivo in newborn Sprague–Dawley rats by injecting 200 μl of blood on postnatal day 4 (P4), and transplanting MSCs (1 × 105 cells) intraventricularly on P6. siRNA-induced BDNF knockdown abolished the in vitro benefits of MSCs on thrombin-induced neuronal cell death. BDNF knockdown also abolished the in vivo protective effects against severe IVH-induced brain injuries such as the attenuation of posthemorrhagic hydrocephalus, impaired behavioral test performance, increased astrogliosis, increased number of TUNEL cells, ED-1+ cells, and inflammatory cytokines, and reduced myelin basic protein expression. Our data indicate that BDNF secreted by transplanted MSCs is one of the critical paracrine factors that play a seminal role in attenuating severe IVH-induced brain injuries in newborn rats.

Key words: Brain-derived neurotrophic factor (BDNF); Intraventricular hemorrhage (IVH); Infant, newborn; Infant, premature; Hydrocephalus; Mesenchymal stem cells (MSCs); Cell transplantation

Received July 4, 2016; final acceptance October 12, 2016. Online prepub date: August 16, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Won Soon Park, M.D., Ph.D., Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, 81 Irwonro, Gangnam-gu, Seoul 06351, South Korea. Tel: +82.2-3410-3523; Fax: +82.2-3410-0043; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jee-Yin Ahn, Ph.D., Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, 2066 SeoburoJangangu, Suwon, Gyeonggido, South Korea. Tel: +82-31-299-6134; Fax: +82-31-299-6139; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 157-170, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
692870
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Intramyocardially Transplanted Neonatal Cardiomyocytes (NCMs) Show Structural and Electrophysiological Maturation and Integration and Dose-Dependently Stabilize Function of Infarcted Rat Hearts

Martina Maass,*1 Benjamin Krausgrill,*1 Simon Eschrig,* Tobias Kaluschke,* Katja Urban,* Gabriel Peinkofer,*† Tobias G. Plenge,* Simon Oeckenpohler,* Martin Raths,* Dennis Ladage,* Marcel Halbach,*† Jurgen Hescheler,† and Jochen Muller-Ehmsen*2

*Department of Internal Medicine III, University Hospital of Cologne, Cologne, Germany
†Institute of Neurophysiology, University of Cologne, Cologne, Germany

Cardiac cell replacement therapy is a promising therapy to improve cardiac function in heart failure. Persistence, structural and functional maturation, and integration of transplanted cardiomyocytes into recipients’ hearts are crucial for a safe and efficient replacement of lost cells. We studied histology, electrophysiology, and quantity of intramyocardially transplanted rat neonatal cardiomyocytes (NCMs) and performed a detailed functional study with repeated invasive (pressure–volume catheter) and noninvasive (echocardiography) analyses of infarcted female rat hearts including pharmacological stress before and 3 weeks after intramyocardial injection of 5 × 106 (low NCM) or 25 × 106 (high NCM) syngeneic male NCMs or medium as placebo (Ctrl). Quantitative real-time polymerase chain reaction (PCR) for Y-chromosome confirmed a fivefold higher persisting male cell number in high  NCM versus low NCM after 3 weeks. Sharp electrode measurements within viable slices of recipient hearts demonstrated that transplanted NCMs integrate into host myocardium and mature to an almost adult phenotype, which might be facilitated through gap junctions between host myocardium and transplanted NCMs as indicated by connexin43 in histology. Ejection fraction of recipient hearts was severely impaired after ligation of left anterior descending (LAD; pressure–volume catheter: 39.2 } 3.6%, echocardiography: 39.9 } 1.4%). Repeated analyses revealed a significant further decline within 3 weeks in Ctrl and a dose-dependent stabilization in cell-treated groups. Consistently, stabilized cardiac function/morphology in cell-treated groups was seen in stroke volume, cardiac output, ventricle length, and wall thickness. Our findings confirm that cardiac cell replacement is a promising therapy for ischemic heart disease since immature cardiomyocytes persist, integrate, and mature after intramyocardial transplantation, and they dose-dependently stabilize cardiac function after myocardial infarction.

Key words: Myocardial infarction (MI); Cell replacement therapy; Neonatal cardiomyocytes (NCMs); Echocardiography; Pressure–volume catheterization

Received April 19, 2016; final acceptance October 14, 2016. Online prepub date: August 17, 2016.
1These authors provided equal contribution to this work.
2Current address: Department of Internal Medicine 3, Asklepios Hospital Altona, Hamburg, Germany.
Address correspondence to Dr. Benjamin Krausgrill, M.D., Department of Internal Medicine III, University Hospital of Cologne, Kerpener Str. 62, 50937 Cologne, Germany. Tel: +49 221 478-87401; Fax: +49 221 478-87372; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it