Cell Transplantation 24(1) Abstracts

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Cell Transplantation, Vol. 24, pp. 1-9, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667493
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Automated Digital Image Analysis of Islet Cell Mass Using Nikon’s Inverted Eclipse Ti Microscope and Software to Improve Engraftment May Help to Advance the Therapeutic Efficacy and Accessibility of Islet Transplantation Across Centers

Valery Gmyr,*Caroline Bonner,*Bruno Lukowiak,*Valerie Pawlowski,*Nathalie Dellaleau,*Sandrine Belaich,*Isanga Aluka,*Ericka Moermann,*Julien Thevenet,*Rimed Ezzouaoui,*Gurvan Queniat,*Francois Pattou,*‡§ and Julie Kerr-Conte*

*European Genomic Institute for Diabetes (EGID), Lille, France
INSERM UMR U859, Lille, France
‡UNIV LILLE 2, Lille, France
§CHULille, Lille, France

Reliable assessment of islet viability, mass, and purity must be met prior to transplanting an islet preparation into patients with type 1 diabetes. The standard method for quantifying human islet preparations is by direct microscopic analysis of dithizone-stained islet samples, but this technique may be susceptible to inter-/intraobserver variability, which may induce false positive/negative islet counts. Here we describe a simple, reliable, automated digital image analysis (ADIA) technique for accurately quantifying islets into total islet number, islet equivalent number (IEQ), and islet purity before islet transplantation. Islets were isolated and purified from n = 42 human pancreata according to the automated method of Ricordi et al. For each preparation, three islet samples were stained with dithizone and expressed as IEQ number. Islets were analyzed manually by microscopy or automatically quantified using Nikon’s inverted Eclipse Ti microscope with built-in NIS-Elements Advanced Research (AR) software. The AIDA method significantly enhanced the number of islet preparations eligible for engraftment compared to the standard manual method (p < 0.001). Comparisons of individual methods showed good correlations between mean values of IEQ number (r2 = 0.91) and total islet number (r2 = 0.88) and thus increased to r2 = 0.93 when islet surface area was estimated comparatively with IEQ number. The ADIA method showed very high intraobserver reproducibility compared to the standard manual method (p < 0.001). However, islet purity was routinely estimated as significantly higher with the manual method versus the ADIA method (p < 0.001). The ADIA method also detected small islets between 10 and 50 μm in size. Automated digital image analysis utilizing the Nikon Instruments software is an unbiased, simple, and reliable teaching tool to comprehensively assess the individual size of each islet cell preparation prior to transplantation. Implementation of this technology to improve engraftment may help to advance the therapeutic efficacy and accessibility of islet transplantation across centers.

Key words: Human islets; Automated digital image analysis (ADIA); Standard manual method; Islet surface area; Transplantation centers

Received November 30, 2012; final acceptance April 18, 2013. Online prepub date: April 29, 2013.
Address correspondence to Julie Kerr-Conte, Ph.D., Univ Lille Nord de France/INSERM U859, Faculty of Medicine, 1 Place de Verdun, F59045 Lille, France. Tel: +33 3 20 62 35 43; Fax: +33 3 20 62 35 40; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 11-23, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X673469
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Impact of Islet Size on Pancreatic Islet Transplantation and Potential Interventions to Improve Outcome

Daria Zorzi,* Tammy Phan,* Marco Sequi,† Yong Lin,* Daniel H. Freeman,‡ Luca Cicalese,* and Cristiana Rastellini*

*Department of Surgery, Texas Transplant Center, University of Texas Medical Branch, Galveston, TX, USA
†Laboratory for Mother and Child Health, Department of Public Health, “Mario Negri” Pharmacological Research Institute, Milan, Italy
‡Department of Epidemiology and Biostatistics, University of Texas Medical Branch, Galveston, TX, USA

Better results have been recently reported in clinical pancreatic islet transplantation (ITX) due mostly to improved isolation techniques and immunosuppression; however, some limitations still exist. It is known that following transplantation, 30% to 60% of the islets are lost. In our study, we have investigated 1) the role of size as a factor affecting islet engraftment and 2) potential procedural manipulations to increase the number of smaller functional islets that can be transplanted. C57/BL10 mice were used as donors and recipients in a syngeneic islet transplant model. Isolated islets were divided by size (large, >300 mm; medium 150–300 mm; small, <150 mm). Each size was transplanted in chemically induced diabetic mice as full (600 IEQ), suboptimal (400 IEQ), and marginal mass (200 IEQ). Control animals received all size islets. Engraftment was defined as reversal of diabetes by day 7 posttransplantation. When the superiority of smaller islets was observed, strategies of overdigestion and fragmentation were adopted during islet isolation in the attempt to reduce islet size and improve engraftment. Smaller islets were significantly superior in engraftment compared to medium, large, and control (all sizes) groups. This was more evident when marginal mass data were compared. In all masses, success decreased as islet size increased. Once islets were engrafted, functionality was not affected by size. When larger islets were fragmented, a significant decrease in islet functionality was observed. On the contrary, if pancreatawere slightly overdigested, although not as successful as small naive islets, an increase in engraftment was observed when compared to the control group. In conclusion, smaller islets are superior in engraftment following islet transplantation. Fragmentation has a deleterious effect on islet engraftment. Islet isolations can be performed by reducing islet size with slight overdigestion, and it can be safely adopted to improve clinical outcome.

Key words: Islet size; Islet engraftment; Islet overdigestion; Islet fragmentation; Islet functionality; Murine model islet transplant

Received March 8, 2013; final acceptance September 12, 2013. Online prepub date: October 18, 2013.
Address correspondence to Cristiana Rastellini, M.D., Professor of Surgery, Medicine, Microbiology and Immunology, Director, Cell Transplant, Director, Transplant Research, University of Texas Medical Branch, 6.312C John Sealy Annex, 301 University Boulevard, Galveston, TX 77555-0533, USA. Tel: +1-409-772-2412; Fax: +1-409-747-7364; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 25-36, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X673441
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Diazoxide, a KATP Channel Opener, Prevents Ischemia–Reperfusion Injury in Rodent Pancreatic Islets

Yong Wang,*1 Shusen Wang,†1 Tricia Harvat,* Katie Kinzer,* Lisa Zhang,* Feng Feng,* Meirigeng Qi,* and Jose Oberholzer*

*University of Illinois at Chicago Department of Transplant/Surgery, Chicago, IL, USA
†Transplant Center, Tianjin First Center Hospital, Tianjin, China

Diazoxide (DZ) is a pharmacological opener of ATP-sensitive K+ channels that has been used for mimicking ischemic preconditioning and shows protection against ischemic damage. Here we investigated whether diazoxide supplementation to University of Wisconsin (UW) solution has cellular protection during islet isolation and improves in vivo islet transplant outcomes in a rodent ischemia model. C57/B6 mice pancreata were flushed with UW or UW+ DZ solution and cold preserved for 6 or 10 h prior to islet isolation. Islet yield, in vitro and in vivo function, mitochondrial morphology, and apoptosis were evaluated. Significantly higher islet yields were observed in the UW+ DZ group than in the UW group (237.5 ± 25.6 vs. 108.7 ± 49.3, p < 0.01). The islets from the UW+ DZ group displayed a significantly higher glucose-induced insulin secretion (0.97 ng/ml ± 0.15 vs. 0.758 ng/ml ± 0.21, p = 0.009) and insulin content (60.96 ng/islet ± 13.94 vs. 42.09 ng/islet ± 8.15, p = 0.002). The DZ-treated islets had well-preserved mitochondrial morphology with superior responses of mitochondrial potentials, and calcium influx responded to glucose. A higher number of living cells and less late apoptotic cells were observed in the UW+ DZ group (p < 0.05). Additionally, the islets from the UW+ DZ group had a significantly higher cure rate and improved glucose tolerance. This study is the first to report mitoprotective effects of DZ for pancreas preservation and islet isolation. In the future, it will be necessary to further understand the underlying mechanism for the mitoprotection and to test this promising approach for pancreas preservation and the islet isolation process in nonhuman primates and ultimately humans.

Key words: Diazoxide (DZ); Cold ischemia; Islet isolation; Islet transplantation; Pancreas preservation; Mitoprotection

Received January 18, 2013; final acceptance September 1, 2013. Online prepub date: September 10, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Yong Wang, University of Illinois at Chicago, Department of Transplant/Surgery, 840 South Wood Street, Clinical Science Building, Suite 402, Chicago, IL 60612, USA. Tel: +1 312.996.0851; Fax: +1 312 413 3483; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or JoseOberholzer, University of Illinois at Chicago, Department of Transplant/Surgery, 840 South Wood Street, Clinical Science Building, Suite 402, Chicago, IL 60612, USA. Tel: +1 312.996.0851; Fax: +1 312 413 3483; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 37-48, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X673423
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Endothelial Progenitor Cells Enhance Islet Engraftment, Influence β-Cell Function, and Modulate Islet Connexin 36 Expression

Daniella Penko,*† Darling Rojas-Canales,*‡ Daisy Mohanasundaram,*‡ Heshan S. Peiris,§ Wai Y. Sun,*¶ Christopher J. Drogemuller,‡ Damien J. Keating,§ P. Toby H. Coates,*†‡ Claudine S. Bonder,*†¶ and Claire F. Jessup*†§

*School of Medicine, Discipline of Medicine, University of Adelaide, Adelaide, SA, Australia
†The Robinson Institute, Centre for Stem Cell Research, University of Adelaide, Adelaide, SA, Australia
‡Central Northern Adelaide Renal and Transplantation Service, Royal Adelaide Hospital, Adelaide, SA, Australia
§Department of Human Physiology and Centre for Neuroscience, School of Medicine, Flinders University of SA, Bedford Park, SA, Australia
¶Vascular Biology and Cell Trafficking Laboratory, Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia

The success of pancreatic islet transplantation is limited by delayed engraftment and suboptimal function in the longer term. Endothelial progenitor cells (EPCs) represent a potential cellular therapy that may improve the engraftment of transplanted pancreatic islets. In addition, EPCs may directly affect the function of pancreatic b-cells. The objective of this study was to examine the ability of EPCs to enhance pancreatic islet transplantation in a murine syngeneic marginal mass transplant model and to examine the mechanisms through which this occurs. We found that cotransplanted EPCs improved the cure rate and initial glycemic control of transplanted islets. Gene expression data indicate that EPCs, or their soluble products, modulate the expression of the b-cell surface molecule connexin 36 and affect glucose-stimulated insulin release in vitro. In conclusion, EPCs are a promising candidate for improving outcomes in islet transplantation, and their mechanisms of action warrant further study.

Key words: Islets of Langerhans; Islet transplantation; Diabetes; Endothelial progenitor cells (EPCs); Connexin 36; Cell therapy

Received December 12, 2012; final acceptance September 1, 2013. Online prepub date: September 10, 2013.
Address correspondence to Dr. Claire F. Jessup, Rm 6E101, Department of Human Physiology, School of Medicine, Flinders University of SA, Bedford Park, SA 5042, Australia. Tel: +61(8) 8204 3960; Fax: +61(8) 8204 5768; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 49-62, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675133
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Adipocyte Regeneration After Free Fat Transplantation: Promotion by Stromal Vascular Fraction Cells

Ming Zhu,*†1 Ziqing Dong,*1 JianHua Gao,* Yunjun Liao,* Jian Xue,† Yi Yuan,* Linqi Liu,* Qiang Chang,* and Feng Lu*

*Department of Plastic and Reconstructive Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China
†Department of Dermatology, Armed Police Hospital, Guangzhou Medical University, Guangzhou, China

Our objective was to explore the mechanism of cell-assisted adipose transplantation by using freshly isolated human stromal vascular fraction (SVF) cells and to observe the dynamic changes of the graft after transplantation. The SVF was isolated from human liposuction aspirates, and 0.5 ml adipose tissue was mixed with 1 × 106 SVF cells or culture medium then injected to nude mice subcutaneously. At 1, 4, 7, 14, 30, 60, and 90 days after transplantation, samples were harvested for 1) general observation and retention rate; 2) whole-mount stain; 3) H&E stain; 4) immunohistochemical staining for S100, CD68, and CD34; 5) ELISA for VEGF and bFGF; 6) peroxisome proliferator-activated receptor-g (PPARg) fluorescence in situ hybridization. The retention rate in the experiment group was markedly higher than that in the control group. Whole-mount stain shows most of the cells in the center of the graft could not survive the ischemia until day 14. Histology showed new vessels on the surface of the graft at 3 days. However, in the control group, fewer newly formed vessels were detected until day 7. In the late stage of transplantation, gradual fibrosis was found in the graft, and the tissue was divided into a grid-like structure. A large number of round neonatal adipocytes with big nuclei in the center were found surrounding the new vessels, which were S100 and CD34 positive and CD68 negative. In the late stage of transplantation, most of the neonatal adipocytes were human PPARg positive. Moreover, the SVF group showed a higher level of VEGF and bFGF. SVF assisting adipose transplantation could increase the retention rate of the graft through promoting adipose tissue regeneration via secretion of growth factors, promotion of angiogenesis, and increasing the density of mesenchymal stem cells.

Key words: Stromal vascular fraction (SVF) cells; Fat transplantation; Cell therapy; Adipocyte regeneration

Received August 7, 2012; final acceptance October 21, 2013. Online prepub date: October 29, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Feng Lu, Department of Plastic and Reconstructive Surgery, Nanfang Hospital, Southern Medical University, 510515 Guangzhou, China. Tel: +0086 20 62787550; Fax: +0086 20 87281445; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jianhua Gao, M.D., Ph.D., Department of Plastic and Reconstructive Surgery, Nanfang Hospital, Southern Medical University, 510515 Guangzhou, China. Tel: +0086 20 62787550; Fax: +0086 20 87281445; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 63-72, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X674062
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Autologous Fat Grafting in the Treatment of Fibrotic Perioral Changes in Patients With Systemic Sclerosis

Nicoletta Del Papa,* Fabio Caviggioli,† Domenico Sambataro,* Eleonora Zaccara,* Valeriano Vinci,‡ Gabriele Di Luca,§ Antonina Parafioriti,¶ Elisabetta Armiraglio,¶ Wanda Maglione,* Riccardo Polosa,# Francesco Klinger,† and Marco Klinger‡

*U.O.C. Day Hospital ReumatologiaOspedale G. Pini, Milano, Italy
†U.O.C. Chirurgia PlasticaMultimedica Holding SpA, Milano, Italy
‡U.O.C. Chirurgia PlasticaIstituto Clinico HumanitasUniversità degli studi di Milano, Milano, Italy
§U.O.S. Chirurgia VascolareOspedale G. Pini, Milano, Italy
¶U.O.C. Anatomia PatologicaOspedale G. Pini, Milano, Italy
#Istituto di Medicina Internad’Emergenza, Dip. Biomedicina ClinicaMolecolareUniversità degli Studi di Catania, Catania, Italy

Autologous fat tissue grafting (AFTG) has been successfully used in the treatment of different sclerotic conditions, including localized scleroderma. Patients with advanced systemic sclerosis (SSc)-related perioral thickening and mouth opening limitation are candidates for this therapeutic approach. AFTG of the lips was performed to improve mouth opening in patients with SSc. We enrolled in the study 20 female patients with diffuse SSc (median age 35 ± 15 years and 11 ± 10 years of disease duration). Two-milliliter fractions of autologous fat drawn from trochanteric or periumbilical areas were injected in eight different sites around the mouth. Baseline and after-treatment mouth opening changes were assessed by measuring interincisal distance and oral perimeter, while skin hardness was tested by digital durometer. Pre- and posttreatment modifications of microvascular architecture were assessed by counting capillaries in the inferior lip videocapillaroscopy (VC) images and by scoring the microvascular density (MVD) in anti-CD34/CD31 immunohistochemical (IH) stained perioral skin biopsy sections. Similarly, histological sections were examined to evaluate dermoepidermic junction (DEJ) modifications. Three months after treatment, both the interincisaldistance and oral perimeter significantly increased (p < 0.001). At the same time, a significant skin neovascularization became evident, both considering the VC images (p < 0.001) and MVD scores in IH sections (p < 0.0001). Finally, some skin histological aspects also improved, as shown by the significant changes in DEJ flattening scores (p < 0.0001). The present study suggests that, in patients with SSc, AFTG can improve mouth opening and function, induce a neovascularization, and partially restore the skin structure.

Key words: Systemic sclerosis (SSc); Adipose tissue; Stem cells; Autologous graft

Received June 25, 2013; final acceptance September 30, 2013. Online prepub date: October 22, 2013.
Address correspondence to Nicoletta Del Papa, M.D., Day Hospital ReumatologiaOspedale G. Pini, Via Pini9 - 20122 Milano, Italy. Tel: +39-0258296663; Fax: +39-0258296495; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 73-83, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674080
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Coculture With Mesenchymal Stem Cells Results in Improved Viability and Function of Human Hepatocytes

Emer Fitzpatrick,* Yue Wu,† Paramjeet Dhadda,† Robin D. Hughes,† Ragai R. Mitry,† Hong Qin,† Sharon C. Lehec,† Nigel D. Heaton,† and Anil Dhawan*

*Paediatric Liver, GI and Nutrition Centre, King’s College London School of Medicine at King’s College Hospital, Denmark Hill, London, UK
†Institute of Liver Studies, King’s College London School of Medicine at King’s College Hospital, Denmark Hill, London, UK

Hepatocyte transplantation is becoming an accepted therapy for acute liver failure, either as a bridge to liver regeneration or to organ transplantation. Hepatocytes provide liver function in place of the failing organ. The maintenance of sufficient viability and function of the transplanted hepatocytes is a concern. There is a lot of recent interest in mesenchymal stem cells (MSCs) for the provision of structural and trophic support to hepatocytes, but few studies currently use primary human hepatocytes. The aim of this study was to investigate if coculture of human MSCs with cryopreserved human hepatocytes may improve their function and viability, thus with potential for cellular therapy of liver disease. MSCs were isolated from human umbilical cord or adipose tissue. Hepatocytes were isolated from donor organs unsuitable for transplantation. MSCs and hepatocytes were cocultured in both direct and indirect contact. Conditioned medium (CM) from cocultured MSCs and hepatocytes was also used on hepatocytes. Viability and liver-specific function were compared between test and controls. Human hepatocytes that were cocultured directly with MSCs demonstrated improved production of albumin from day 5 to day 25 of culture. This effect was most prominent at day 15. Likewise, urea production was improved in coculture from day 5 to 25. Indirect coculture demonstrated improved albumin production by day 4 (1,107 ng/ml) versus hepatocyte monoculture (940 ng/ml). Hepatocytes in CM demonstrated a nonsignificantimprovement in function. The viability of cocultured hepatocytes was superior to that of monocultured cells with up to a 16% improvement. Thus, coculture of human hepatocytes with MSCs demonstrates both improved function and viability. The effect is seen mainly with direct coculture but can also be seen in indirect culture and with CM. Such coculture conditions may convey major advantages in hepatocyte survival and function for cell transplantation.

Key words: Mesenchymal stem cells (MSCs); Hepatocyte transplantation; Acute liver failure

Received January 31, 2013; final acceptance October 4, 2013. Online prepub date: October 18, 2013.
Address correspondence to Emer Fitzpatrick, Paediatric Liver GI and Nutrition Centre, King’s College Hospital, Denmark Hill, London SE5 9PJ, UK. Tel: +44 2032994643; Fax: +44 2032994228; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 85-95, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667736
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Bone Marrow-Derived c-kit+ Cells Attenuate Neonatal Hyperoxia-Induced Lung Injury

Shalini Ramachandran,*† Cleide Suguihara,*† Shelley Drummond,*† Konstantinos Chatzistergos,‡§ Jammie Klim,* Eneida Torres,*† Jian Huang,*† Dorothy Hehre,*† Claudia O. Rodrigues,§¶ Ian K.McNiece,‡§ Joshua M. Hare,‡§ and Karen C. Young*†§

*Department of Pediatrics/Division of Neonatology, University of Miami Miller School of Medicine, Miami, FL, USA
Batchelor Children’s Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA
‡Department of Medicine/Cardiovascular Division, University of Miami Miller School of Medicine, Miami, FL, USA
§The Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, USA
¶Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA

Recent studies suggest that bone marrow (BM)-derived stem cells have therapeutic efficacy in neonatal hyperoxia-induced lung injury (HILI). c-kit, a tyrosine kinase receptor that regulates angiogenesis, is expressed on several populations of BM-derived cells. Preterm infants exposed to hyperoxia have decreased lung angiogenesis. Here we tested the hypothesis that administration of BM-derived c-kit+ cells would improve angiogenesis in neonatal rats with HILI. To determine whether intratracheal (IT) administration of BM-derived c-kit+ cells attenuates neonatal HILI, rat pups exposed to either normobaric normoxia (21% O2) or hyperoxia (90% O2) from postnatal day (P) 2 to P15 were randomly assigned to receive either IT BM-derived green fluorescent protein (GFP)+ c-kit cells (PL) or BM-derived GFP+ c-kit+ cells on P8. The effect of cell therapy on lung angiogenesis, alveolarization, pulmonary hypertension, vascular remodeling, cell proliferation, and apoptosis was determined at P15. Cell engraftment was determined by GFP immunostaining. Compared to PL, the IT administration of BM-derived c-kit+ cells to neonatal rodents with HILI improved alveolarization as evidenced by increased lung septation and decreased mean linear intercept. This was accompanied by an increase in lung vascular density, a decrease in lung apoptosis, and an increase in the secretion of proangiogenic factors. There was no difference in pulmonary vascular remodeling or the degree of pulmonary hypertension. Confocal microscopy demonstrated that 1% of total lung cells were GFP+ cells. IT administration of BM-derived c-kit+cells improves lung alveolarization and angiogenesis in neonatal HILI, and this may be secondary to an improvement in the lung angiogenic milieu.

Key words: c-kit; Hyperoxia; Stem cells; Angiogenesis; Bronchopulmonary dysplasia

Received December 18, 2012; final acceptance May 13, 2013. Online prepub date: May 22, 2013.
Address correspondence to Karen C. Young, M.D., University of Miami Miller School of Medicine, Batchelor Children’s Research Institute, 1580 NW 10th Avenue RM-344, Miami, FL 33136, USA. Tel: +1-305-243-4531; Fax: +1-305-243-6114; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 97-113, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674675
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Mouse Dental Pulp Stem Cells Support Human Umbilical Cord Blood-Derived Hematopoietic Stem/Progenitor Cells In Vitro

Ryusuke Nakatsuka,* Yoshikazu Matsuoka,* Yasushi Uemura,† Keisuke Sumide,* Ryuji Iwaki,* Masaya Takahashi,* Tatsuya Fujioka,* Yutaka Sasaki,* and Yoshiaki Sonoda*

*Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, Hirakata, Osaka, Japan
†Division of Cancer Immunotherapy, Exploratory Oncology Research and Clinical Trial Center National Cancer Center, Kashiwa, Chiba, Japan

It is well documented that specialized mesenchymal stem/stromal cells (MSCs) constitute the hematopoietic stem cell (HSC) niche in the bone marrow (BM), and these MSCs support/maintain the HSCs in an undifferentiated state. A number of studies have demonstrated that BM-derived MSCs (BM-MSCs) can support HSCs in vitro. However, it remains unclear whether nonhematopoietic tissue-derived MSC-like cells, such as dental pulp stem cells (DPSCs), have the ability to support HSCs. In this study, we prospectively isolated DPSCs from mouse mandibular incisors by fluorescence-activated cell sorting (FACS) using BM-MSC markers, such as PDGFRa and Sca-1. The PDGFRa and Sca-1 double-positive DPSCs and BM-MSCs showed similar morphologies and expression patterns of MSC markers. The ability of the DPSCs to support hematopoietic stem/progenitor cells (HSPCs) was then analyzed by an in vitro coculture system. Moreover, their HSC-supporting activity was evaluated by in vivo xenotransplantation assays using NOD/Shi-scid/IL-2Rgcnull (NOG) mice. Interestingly, the DPSCs supported human cord blood (CB)-derived CD34-positive (CD34+), as well as CD34-negative (CD34), HSCs. The supporting activities of DPSCs for human CB-derived CD34+ and CD34 HSCs were comparable to those of BM-MSCs. The results of the present study demonstrated, for the first time, that prospectively isolated murine PDGFRa and Sca-1 double-positive DPSCs could support primitive human CD34+ and CD34 HSCs in vitro.

Key words: Dental pulp stem cells (DPSCs); Cord blood; Xenotransplantation; Hematopoietic stem/progenitor cells; SCID-repopulating cells; Mesenchymal stem/stromal cells

Received September 11, 2013; final acceptance October 23, 2013. Online prepub date: October 29, 2013.
Address correspondence to Yoshiaki Sonoda, M.D., Ph.D., Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, 2-5-1 Shin-machi, Hirakata, Osaka 570-8506, Japan. Tel: +81-72-804-2391; Fax: +81-72-804-2399; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 115-131, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674657
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Permissive Schwann Cell Graft/Spinal Cord Interfaces for Axon Regeneration

Ryan R. Williams,* Martha Henao,* Damien D. Pearse,*† and Mary Bartlett Bunge*†‡

*The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA
†Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, USA
‡Department of Cell Biology, University of Miami Miller School of Medicine, Miami, FL, USA

The transplantation of autologous Schwann cells (SCs) to repair the injured spinal cord is currently being evaluated in a clinical trial. In support, this study determined properties of spinal cord/SC bridge interfaces that enabled regenerated brainstem axons to cross them, possibly leading to improvement in rat hindlimb movement. Fluid bridges of SCs and Matrigel were placed in complete spinal cord transections. Compared to pregelled bridges of SCs and Matrigel, they improved regeneration of brainstem axons across the rostral interface. The regenerating brainstem axons formed synaptophysin+ bouton-like terminals and contacted MAP2A+ dendrites at the caudal interface. Brainstem axon regeneration was directly associated with glial fibrillary acidic protein (GFAP+) astrocyte processes that elongated into the SC bridge. Electron microscopy revealed that axons, SCs, and astrocytes were enclosed together within tunnels bounded by a continuous basal lamina. Neuroglycan (NG2) expression was associated with these tunnels. One week after injury, the GFAP+ processes coexpressed nestin and brain lipid-binding protein, and the tips of GFAP+/NG2+ processes extended into the bridges together with the regenerating brainstem axons. Both brainstem axon regeneration and number of GFAP+ processes in the bridges correlated with improvement in hindlimb locomotion. Following SCI, astrocytes may enter a reactive state that prohibits axon regeneration. Elongation of astrocyte processes into SC bridges, however, and formation of NG2+ tunnels enable brainstem axon regeneration and improvement in function. It is important for spinal cord repair to define conditions that favor elongation of astrocytes into lesions/transplants.

Key words: Spinal cord injury; Schwann cells (SCs); Astrocytes; Neuroglycan (NG2); Axon regeneration; Transplantation; Interface

Received May 20, 2013; final acceptance October 9, 2013. Online prepub date: October 22, 2013.
Address correspondence to Mary Bartlett Bunge, Ph.D., The Miami Project to Cure Paralysis, Lois Pope Life Center, PO Box 016960, R-48, Miami, FL, 33101, USA. Tel: +1-305-243-4596; Fax: +1-305-243-3923; E-mail: MBunge This e-mail address is being protected from spambots. You need JavaScript enabled to view it