Cell Transplantation 25(6) Abstracts

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Cell Transplantation, Vol. 25, pp. 995-1005, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688560
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Review

Hematopoietic Cell and Renal Transplantation in Plasma Cell Dyscrasia Patients

Olga Baraldi, Valeria Grandinetti, Gabriele DonatiGiorgia Comai, Giuseppe Battaglino, Vania Cuna, Irene Capelli, Elisa Sala, and Gaetano La Manna

Department of Experimental Diagnostic and Specialty Medicine (DIMES), Nephrology, Dialysis and Renal Transplant Unit, St Orsola Hospital, University of Bologna, Bologna, Italy

Gammopathies, multiple myeloma, and amyloidosis are plasma dyscrasias characterized by clonal proliferation and immunoglobulin overproduction. Renal impairment is the most common and serious complication with an incidence of 20–30% patients at the diagnosis. Kidney transplant has not been considered feasible in the presence of plasma dyscrasias because the immunosuppressive therapy may increase the risk of neoplasia progression, and paraproteins may affect the graft. However, recent advances in clinical management of multiple myeloma and other gammopathies allow considering kidney transplant as a possible alternative to dialysis. Numerous evidence indicates the direct relationship between hematological remission and renal function restoring. The combination of kidney and hematopoietic cell transplant has been reported as a promising approach to reestablish end-organ function and effectively treat the underlying disease. This review describes current protocols used to perform kidney transplantation in patients with plasma dyscrasias.

Key words: Plasma dyscrasia; Multiple myeloma (MM); Amyloidosis (AL); Hematopoietic cell transplantation; Kidney transplant

Received June 27, 2014; final acceptance July 3, 2015. Online prepub date: July 8, 2015.
Address correspondence to Gaetano La Manna, M.D. Ph.D., Policlinico Universitario S. Orsola Malpighi, Pad 15, Via Massarenti, 9, 40128 – Bologna, Università di Bologna, Bologna, Italy. Tel: +39 051 6364577; Fax +39 051 344439; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



Cell Transplantation, Vol. 25, pp. 1007-1023, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688632
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Review

Pluripotent Stem Cells to Rebuild a Kidney: The Lymph Node as a Possible Developmental Niche

Maria Giovanna Francipane*† and Eric Lagasse*

*McGowan Institute for Regenerative Medicine, Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
Ri.MED Foundation, Palermo, Italy

Kidney disease poses a global challenge. Stem cell therapy may offer an alternative therapeutic approach to kidney transplantation, which is often hampered by the limited supply of donor organs. While specific surface antigen markers have yet to be identified for the analysis and purification of kidney stem/progenitor cells for research or clinical use, the reprogramming of somatic cells to pluripotent cells and their differentiation into the various kidney lineages might represent a valuable strategy to create a renewable cell source for regenerative purposes. In this review, we first provide an overview of kidney development and explore current knowledge about the role of extra- and intrarenal cells in kidney repair and organogenesis. We then discuss recent advances in the 1) differentiation of rodent and human embryonic stem cells (ESCs) into renal lineages; 2) generation of induced pluripotent stem cells (iPSCs) from renal or nonrenal (kidney patient-derived) adult cells; 3) differentiation of iPSCs into renal lineages; and 4) direct transcriptional reprogramming of adult renal cells into kidney progenitor cells. Finally, we describe the lymph node as a potential three-dimensional (3D) in vivo environment for kidney organogenesis from pluripotent stem cells.

Key words: Kidney; Pluripotent stem cells (PSCs); Cellular reprogramming; Lymph node

Received March 31, 2015; final acceptance July 14, 2015. Online prepub date: July 8, 2015.
Address correspondence to Eric Lagasse, Pharm.D., Ph.D., 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA. Tel: 412-624-5285; Fax: 412-624-5363; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Maria Giovanna Francipane, Ph.D., 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1025-1042, 2016
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DOI: http://dx.doi.org/10.3727/096368915X689703
E-ISSN 1555-3892
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The Effect of Fetal Bovine Serum (FBS) on Efficacy of Cellular Reprogramming for Induced Pluripotent Stem Cell (iPSC) Generation

Daekee Kwon,*1 Jin-Su Kim,*1 Byung-Hyun Cha,* Kwang-Sook Park,* Inbo Han,† Kyung-Soon Park,* Hojae Bae,‡ Myung-Kwan Han,§ Kwang-Soo Kim,¶ and Soo-Hong Lee*

*Department of Biomedical Science, College of Life Science, CHA University, Gyeonggi-do, Republic of Korea
†Department of OrthopaedicsBundang CHA Hospital, Sungnam, Republic of Korea
‡College of Animal Bioscience and Technology, Department of Bioindustrial Technologies, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, Republic of Korea
§Departments of Microbiology and Biochemistry, Chonbuk National University, Jeonju, Republic of Korea
¶Molecular Neurobiology Laboratory, McLean Hospital, Harvard Medical School, Belmont, MA, USA

Induced pluripotent stem cells (iPSCs) are pivotal to the advancement of regenerative medicine. However, the low efficacy of iPSC generation and insufficient knowledge about the reprogramming mechanisms involved in somatic cell/adult stem cell reversion to a pluripotent phenotype remain critical hurdles to the therapeutic application of iPSCs. The present study investigated whether the concentration of fetal bovine serum (FBS), a widely employed cell culture additive, can influence the cellular reprogramming efficacy (RE) of human adipose-derived stem cells (hADSCs) to generate iPSCs. Compared with the typically employed concentration of FBS (10%), high concentrations (20% and 30%) increased the RE of hADSCs by approximately twofold, whereas a low concentration (5%) decreased the RE by the same extent. Furthermore, cell counting kit-8 (CCK-8), bromodeoxyuridine (BrdU) incorporation, and fluorescence-activated cell sorting (FACS) assays showed that hADSC proliferation during reprogramming was significantly enhanced by FBS at 20% and 30%, whereas quantitative polymerase chain reaction (qPCR) and Western blotting assays revealed a concomitant decrease in p53, p51, and p21 expression. In addition, the efficacy of retrovirus-mediated transduction into hADSCs was increased by approximately 10% at high concentrations of FBS. It was confirmed that platelet-derived growth factor in the FBS enhanced proliferation and reprogramming efficacy. Finally, the generated iPSCs showed a normal karyotype, the same fingerprinting pattern as parental hADSCs, a genome-wide transcriptome pattern similar to that of human embryonic stem cells (hESCs), and in vivo pluripotency. In conclusion, the current investigation demonstrated that high concentrations of FBS can modulate molecular and cellular mechanisms underlying the reprogramming process in hADSCs, thereby augmenting the cellular RE for iPSC generation.

Key words: Concentration of fetal bovine serum (FBS); Reprogramming efficacy (RE); Proliferation; p53; Transduction efficacy

Received July 3, 2014; final acceptance February 18, 2016. Online prepub date: October 7, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Soo-Hong Lee, Ph.D., Department of Biomedical Science, CHA University, 502 Yatap-dong, Bundang-guSeongnam-siGyeonggi-do 463-840, Republic of Korea. Tel: +82-31-8017-9415; Fax: +82-31-8017-9892; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1043-1056, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689514
E-ISSN 1555-3892
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Human Lipoaspirate as Autologous Injectable Active Scaffold for One-Step Repair of Cartilage Defects

Michela Bosetti,* Alessia Borrone,† Antonia Follenzi,† Fanuel Messaggio,‡ Carlo Tremolada,§ and Mario Cannas†

*Dipartimento di Scienze del FarmacoUniversita del Piemonte Orientale, Novara, Italy
Dipartimento di Scienze della Salute, Universita del Piemonte Orientale, Novara, Italy
‡Diabetes Research Institute, University of Miami, Miami, FL, USA
§IMAGE Institute, Milan, Italy

Research on mesenchymal stem cells from adipose tissue shows promising results for cell-based therapy in cartilage lesions. In these studies, cells have been isolated, expanded, and differentiated in vitro before transplantation into the damaged cartilage or onto materials used as scaffolds to deliver cells to the impaired area. The present study employed in vitro assays to investigate the potential of intra-articular injection of microfragmented lipoaspirate as a one-step repair strategy; it aimed to determine whether adipose tissue can act as a scaffold for cells naturally present at their anatomical site. Cultured clusters of lipoaspirate showed a spontaneous outgrowth of cells with a mesenchymal phenotype and with multilineage differentiation potential. Transduction oflipoaspirate clusters by lentiviral vectors expressing GFP evidenced the propensity of the outgrown cells to repopulate fragments of damaged cartilage. On the basis of the results, which showed an induction of proliferation and ECM production of human primary chondrocytes, it was hypothesized that lipoaspirate may play a paracrine role. Moreover, the structure of a floating culture of lipoaspirate, treated for 3 weeks with chondrogenic growth factors, changed: tissue with a high fat component was replaced by a tissue with a lower fat component and connective tissue rich in GAG and in collagen type I, increasing the mechanical strength of the tissue. From these promising in vitro results, it may be speculated that an injectable autologous biologically active scaffold (lipoaspirate), employed intra-articularly, may 1) become a fibrous tissue that provides mechanical support for the load on the damaged cartilage; 2) induce host chondrocytes to proliferate and produce ECM; and 3) provide cells at the site of injury, which could regenerate or repair the damaged or missing cartilage.

Key words: Lipoaspirate; Cartilage defects; Injectable autologous active scaffold; In vitro

Received March 17, 2015; final acceptance December 18, 2015. Online prepub date: September 21, 2015.
Address correspondence to Michela Bosetti, Ph.D., Dipartimento di Scienze del FarmacoUniversita del Piemonte Orientale, Laboratorio Anatomia Umana, Via Solaroli 17, 28100 Novara, Italia. Tel: +39-0321-660557; Fax: +39-0321-660633; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1057-1072, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687949
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Treatment of Collagen-Induced Arthritis Using Immune Modulatory Properties of Human Mesenchymal Stem Cells

Kyu-Hyung Park, Chin Hee MunMi-Il Kang, Sang-Won Lee, Soo-Kon Lee, and Yong-Beom Park

Division of Rheumatology, Department of Internal Medicine, Institute for Immunology and Immunological Disease, Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, Korea

Mesenchymal stem cells (MSCs) have immune modulatory properties. We investigated the potential therapeutic effects of human bone marrow (BM)-, adipose tissue (AD)-, and cord blood (CB)-derived MSCs in an experimental animal model of rheumatoid arthritis (RA) and explored the mechanism underlying immune modulation by MSCs. We evaluated the therapeutic effect of clinically available human BM-, AD-, and CB-derived MSCs in DBA/1 mice with collagen-induced arthritis (CIA). CIA mice were injected intraperitoneally with three types of MSCs. Treatment control animals were injected with 35 mg/kg methotrexate (MTX) twice weekly. Clinical activity in CIA mice, degree of inflammation, cytokine expression in the joint, serum cytokine levels, and regulatory T cells (Tregs) were evaluated. Mice treated with human BM-, AD-, and CB-MSCs showed significant improvement in clinical joint score, comparable to MTX-treated mice. Histologic examination showed greatly reduced joint inflammation and damage in MSC-treated mice compared with untreated mice. Microcomputed tomography also showed little joint damage in the MSC-treated group. MSCs significantly decreased serum interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, and interferon-γ and increased IL-10 and transforming growth factor-β levels. Tregs were increased in mice treated with MSCs compared to untreated or MTX-treated mice. Human BM-, AD-, and CB-MSCs significantly suppressed joint inflammation in CIA mice. The cells decreased proinflammatory cytokines and upregulated anti-inflammatory cytokines and induced Tregs. Therefore, our study suggests that the use of human BM-, AD-, and CB-MSCs could be an effective therapeutic approach for RA.

Key words: Rheumatoid arthritis (RA); Mesenchymal stem cells (MSCs); Immune modulation; Regulatory T cells (Tregs)

Received November 4, 2013; final acceptance March 31, 2015. Online prepub date: April 7, 2015.
Address correspondence to Yong-Beom Park, M.D., Ph.D., Division of Rheumatology, Department of Internal Medicine, Institute for Immunology and Immunological Disease, and Department of Medical Sciences, Yonsei University College of Medicine, 50 Yonsei-roSeodaemun-gu, Seoul 120-752, South Korea. Tel: +82-2-2228-1967; Fax: +82-2-393-5420; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1073-1083, 2016
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DOI: http://dx.doi.org/10.3727/096368915X689613
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Cell Magnetic Targeting System for Repair of Severe Chronic Osteochondral Defect in a Rabbit Model

Elhussein Elbadry Mahmoud,*† Goki Kamei,* Yohei Harada,* Ryo Shimizu,* Naosuke Kamei,* Nobuo Adachi,* Nabil Ahmed Misk,‡ and Mitsuo Ochi*

*Department of Orthopaedic Surgery, Integrated Health Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
†Department of Surgery, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt
‡Department of Surgery, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt

The aim of this study was to investigate a cell delivery system for repair of severe chronic osteochondral defects using magnetically labeled mesenchymal stem cells (m-MSCs), with the aid of an external magnetic device, through the accumulation of a small number of m-MSCs into a desired area and to detect the suitable number of autologous m-MSCs needed for repair of the defect. Twenty-six male Japanese white rabbits aged 6 months were used. An osteochondral defect was created bilaterally at the weight-bearing surface of the medial femoral condyle of the rabbits’ knees (3 mm diameter; 4 mm depth). At 4 weeks after creation of the defect, autogenic transplantation of the m-MSCs into the defect area was performed, followed by 10-min exposure to an external magnetic device, where animals were divided into four groups: high (1 × 106
m-MSCs), medium (2 × 105 m-MSCs), low (4 × 104 m-MSCs), and control (PBS injection). At 4 and 12 weeks posttransplantation of m-MSCs, repaired tissue was assessed histologically using the Fortier score with toluidine blue staining. Transplantation of a low number of m-MSCs was not enough to improve osteogenesis and chondrogenesis, but the medium and high groups improved repair of the chronic defect withchondrogenic tissues and showed histologically significantly better results than the control and low groups. The use of a magnetic targeting system for delivering m-MSCs has the potential to overcome the clinical hurdles for repair of the severe chronic osteochondral defect. Furthermore, this system is predicted to produce good clinical outcomes for humans, not only to repair osteochondral defects but also to repair a variety of damaged tissues.

Key words: Osteochondral repair; Magnetically labeled mesenchymal stem cells (m-MSCs); Cell delivery system; External magnetic device; Clinical relevance

Received January 6, 2015; final acceptance February 17, 2016. Online prepub date: September 28, 2015.
Address correspondence to Prof./Dr. Mitsuo Ochi, M.D., Ph.D., Department of Orthopaedic Surgery, Integrated Health Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. Tel: +81 82 257 5233; Fax: +81 82 257 5234; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1085-1099, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689550
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Cell-Based Therapy in TBI: Magnetic Retention of Neural Stem Cells In Vivo

Wei-Bin Shen,*1 Céline Plachez,*1,2 Orest Tsymbalyuk,† Natalya Tsymbalyuk,† Su Xu,‡ Aaron M. Smith,§ Sarah L. J. Michel,§ Deborah Yarnell,¶ Roger Mullins,‡ Rao P. Gullapalli,‡ Adam Puche,# J. Marc Simard,**‡‡ Paul S. Fishman,††‡‡ and Paul Yarowsky

*Department of Pharmacology, University of Maryland School of Medicine, Baltimore, MD, USA
†Department of Neurosurgery, University of Maryland School of Medicine, Baltimore, MD, USA
‡Department of Diagnostic Radiology, Nuclear Medicine, University of Maryland School of Medicine, Baltimore, MD, USA
§Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD, USA
¶Research Service, VA Maryland Healthcare System, Baltimore, MD, USA
#Department of Anatomy and Neurobiology, University of Maryland School of Medicine, Baltimore, MD, USA
**Department of Neurosurgery, University of Maryland School of Medicine, Baltimore, MD, USA
††Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, USA
‡‡VA Maryland Healthcare System, Baltimore, MD, USA

Stem cell therapy is under active investigation for traumatic brain injury (TBI). Noninvasive stem cell delivery is the preferred method, but retention of stem cells at the site of injury in TBI has proven challenging and impacts effectiveness. To investigate the effects of applying a magnetic field on cell homing and retention, we delivered human neuroprogenitor cells (hNPCs) labeled with a superparamagnetic nanoparticle into post-TBI animals in the presence of a static magnetic field. We have previously devised a method of loading hNPCs with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles Molday ION Rhodamine B (MIRB™). Labeling of hNPCs (MIRB-hNPCs) does not affect hNPC viability, proliferation, or differentiation. The 0.6 tesla (T) permanent magnet was placed ~4 mm above the injured parietal cortex prior to intracarotid injection of 4 × 104
MIRB-hNPCs. Fluorescence imaging, Perls’ Prussian blue histochemistry, immunocytochemistry with SC121, a human-specific antibody, and T2-weighted magnetic resonance imaging ex vivo revealed there was increased homing and retention of MIRB-hNPCs in the injured cortex as compared to the control group in which MIRB-hNPCs were injected in the absence of a static magnetic field. Fluoro-Jade C staining and immunolabeling with specific markers confirmed the viability status of MIRB-hNPCs posttransplantation. These results show that increased homing and retention of MIRB-hNPCs post-TBI by applying a static magnetic field is a promising technique to deliver cells into the CNS for treatment of neurological injuries and neurodegenerative diseases.

Key words: Traumatic brain injury rat model; Stem cell transplantation; Magnetic stem cell retention; Human neuroprogenitor cells (hNPCs); Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles

Received February 27, 2015; final acceptance November 10, 2015. Online prepub date: September 21, 2015.
1These authors provided equal contribution to this work.
2Current affiliation: The Hussman Institute for Autism, Baltimore, MD, USA.
Address corresponding to Paul Yarowsky, Ph.D., University of Maryland School of Medicine, Department of Pharmacology, 655 W. Baltimore St., Rm 4-002 Bressler Research Bldg., Baltimore, MD 21201, USA. Tel: +1-410-706-3134; Fax: +1-410-706-0032; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1101-1113, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689938
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Combined MSC-Secreted Factors and Neural Stem Cell Transplantation Promote Functional Recovery of PD Rats

Yuan Yao,*1 Chen Huang,*1 Ping Gu,† and Tieqiao Wen*

*Laboratory of Molecular Neural Biology, School of Life Sciences, Shanghai University, Shanghai, China
†Department of Ophthalmology, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China

Stem cell transplantation has enormous potential for the treatment of neurodegenerative disorders like Parkinson’s disease (PD). Mesenchymal stem cells (MSCs) have attracted much attention because they can secrete a wide variety of cellular factors that promote cell growth. In this study, we prepared a conditioned medium (CM) using lyophilized MSC culture medium that contained the secretome of MSCs and applied this CM to the culture of neural stem cells (CM-NSCs) for the transplantation of PD model rats. Quantitative real-time PCR, Western blot, and immunocytochemistry were used to identify cell differentiation and expression of dopaminergic neuron-specific genes in vitro. Behavioral tests including rotational behavior and MWM training tests were also performed to assess the recovery. Our results indicated that combined treatment of CM and neural stem cell transplantation can significantly reduce apomorphine-induced rotational asymmetry and improve spatial learning ability. The CM-NSCs were able to differentiate into dopaminergic neurons in the ventral tegmental area (VTA) and medial forebrain bundle (MFB), and migrated around the lesion site. They showed a higher activity than untreated NSCs in cell survival, migration, and behavior improvement in the dopa-deficit rat model. These findings suggest that the neural stem cells treated with conditioned medium possess a great potential as a graft candidate for the treatment of Parkinson’s disease.

Key words: Secreted factor; Mesenchymal stem cells (MSCs); Neural stem cells (NSCs); Parkinson’s disease (PD); Transplantation

Received December 12, 2014; final acceptance February 25, 2016. Online prepub date: November 24, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Tieqiao Wen, School of Life Sciences, Shanghai University, 333 Nanchen Road, Shanghai, 200444, China. Tel/Fax: 86-021-66132512; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1115-1130, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689910
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Comparative Study on the Differentiation of Mesenchymal Stem Cells Between Fetal and Postnatal Rat Spinal Cord Niche

Songying Cao, Xiaowei Wei, Hui Li, Jianing Miao, Guifeng Zhao, Di Wu, Bo Liu, Yi Zhang, Hui Gu, Lili Wang, Yang Fan, Dong An, and Zhengwei Yuan

Department of Pediatric Surgery, Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang, China

In a previous study, we established a prenatal surgical approach and transplanted mesenchymal stem cells (MSCs) into the fetal rat spinal column to treat neural tube defects (NTDs). We found that the transplanted MSCs survived and differentiated into neural lineage cells. Various cytokines and extracellular signaling systems in the spinal cord niche play an important role in cell differentiation. In this study, we observed the differentiation of transplanted MSCs in different spinal cord niches and further observed the expression of neurotrophic factors and growth factors in the spinal cord at different developmental stages to explore the mechanism of MSC differentiation in different spinal cord niches. The results showed that transplanted MSCs expressed markers of neural precursor cells (nestin), neurogliocytes (GFAP), and neurons (β-tubulin). The percentages of GFP+/nestin+
double-positive cells in transplanted MSCs in E16, P1, and P21 rats were 18.31%, 12.18%, and 5.06%, respectively. The percentages of GFP+/GFAP+double-positive cells in E16, P1, and P21 rats were 32.01%, 15.35%, and 12.56%, respectively. The percentages of GFP+/β-tubulin+ double-positive cells in E16, P1, and P21 were 11.76%, 7.62%, and 4.88%, respectively. The differentiation rates of MSCs in embryonic spinal cords were significantly higher than in postnatal spinal cords (p < 0.05). We found that the transplanted MSCs expressed synapsin-1 at different developmental stages. After MSC transplantation, we observed that neurotrophic factor-3 (NT-3), fibroblast growth factor-2 (FGF-2), FGF-8, transforming growth factor-α (TGF-α), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) significantly increased in the MSC transplantation group compared with the blank injection group. Furthermore, FGF-2 and VEGF expression were positively correlated with the number of surviving MSCs. In addition, we found that the expression of brain-derived neurotrophic factor (BDNF), NT-3, FGF-8, TGF-β, epidermal growth factor (EGF), and insulin-like growth factor (IGF) decreased with age, and the expression of FGF-2, FGF-10, FGF-20, TGF-α, and PDGF increased with age. Our data suggest that the embryonic spinal cord niche is more conducive to MSC differentiation after transplantation.

Key words: Mesenchymal stem cells (MSCs); Niche; Cell transplantation; Neurotrophic factors; Growth factors

Received October 11, 2014; final acceptance February 23, 2016. Online prepub date: November 19, 2015.
Address correspondence to Dr. Zhengwei Yuan, Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, No. 36, Sanhao Street, Heping District, Shenyang, Liaoning Province, 110004, P. R. China. Tel: +86 24 23929903; Fax: +86 24 23929903; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1131-1144, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689640
E-ISSN 1555-3892
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Optimal Timing of Mesenchymal Stem Cell Therapy for Neonatal Intraventricular Hemorrhage

Won Soon Park,*1 Se In Sung,*1 So Yoon Ahn,* Dong Kyung Sung,† Geun Ho Im,† Hye Soo Yoo,* Soo Jin Choi,‡ and Yun Sil Chang*

*Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
†Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Seoul, Korea
‡Biomedical Research Institute, MEDIPOST Co., Ltd., Seoul, Korea

We recently showed that intraventricular transplantation of human umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) significantly attenuated posthemorrhagic hydrocephalus (PHH) and brain injury after severe intraventricular hemorrhage (IVH) in newborn rat pups. The purpose of this study was to optimize the timing of MSC transplantation for severe IVH. Severe IVH was induced by injecting 100 μl of blood into each ventricle of Sprague–Dawley rats on postnatal day 4 (P4). Human UCB-derived MSCs (1 × 105
cells in 10 μl of normal saline) were transplanted intraventricularly under stereotaxic guidance either early at P6 or late at P11. Serial brain MRIs and behavioral function tests, such as negative geotaxis and rotarod tests, were performed. At P32, brain tissue samples were obtained for histological and biochemical analyses. Intracerebroventricular transplantation of MSCs significantly attenuated the development of PHH, behavioral impairment, increased apoptosis and astrogliosis, reduced corpus callosum thickness and brain myelination, and upregulated inflammatory cytokines including interleukin (IL)-1α, IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) at P6 but not at P11 after induction of severe IVH. Intracerebroventriculartransplantation of human UCB-derived MSCs attenuated PHH and brain injury after severe IVH in newborn rats in a time-dependent manner. Significant neuroprotection was only demonstrated when administered early at 2 days after induction but not late at 7 days after induction of severe IVH.

Key words: Intraventricular hemorrhage (IVH); Infant; Newborn; Mesenchymal stem cells (MSCs); Cell transplantation

Received June 25, 2014; final acceptance November 25, 2015. Online prepub date: October 5, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Yun Sil Chang, M.D., Ph.D., Department of Pediatrics, Samsung Medical Center; Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul 135-710, Korea. Tel: +82-2-3410-3528; 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


Cell Transplantation, Vol. 25, pp. 1145-1157, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688263
E-ISSN 1555-3892
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Retinal Angiogenesis Effects of TGF-β1 and Paracrine Factors Secreted From Human Placental Stem Cells in Response to a Pathological Environment

Kyung-Sul Kim,*1 Ji-Min Park,*†1 TaeHo Kong,*† Chul Kim,*† Sang-hun Bae,*† Han Wool Kim,† and Jisook Moon*

*College of Life Science, Department of Biotechnology, CHA University, Seoul, Korea
†General Research Institute, Gangnam CHA General Hospital, Seoul, Korea

Abnormal angiogenesis is a primary cause of many eye diseases, including diabetic retinopathy, age-related macular degeneration, and retinopathy of prematurity. Mesenchymal stem cells (MSCs) are currently being investigated as a treatment for several such retinal diseases based on their neuroprotective and angiogenic potentials. In this study, we evaluated the role of systemically injected human placental amniotic membrane-derived MSCs (AMSCs) on pathological neovascularization of proliferative retinopathy. We determined that AMSCs secrete higher levels of transforming growth factor-β (TGF-β1) than other MSCs, and the secreted TGF-b1 directly suppresses the proliferation of endothelial cells under pathological conditions in vitro. Moreover, in a mouse model of oxygen-induced retinopathy, intraperitoneally injected AMSCs migrated into the retina and suppressed excessive neovascularization of the vasculature via expression of TGF-β1, and the antineovascular effect of AMSCs was blocked by treatment with TGF-β1 siRNA. These findings are the first to demonstrate that TGF-β1 secreted from AMSCs is one of the key factors to suppress retinal neovascularization in proliferative retinopathy and further elucidate the therapeutic function of AMSCs for the treatment of retinal neovascular diseases.

Key words: Retina; Angiogenesis; Placenta; Mesenchymal stem cells (MSCs); Systemic delivery; Transforming growth factor-β (TGF-β1)

Received December 22, 2013; final acceptance September 18, 2015. Online prepub date: June 10, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Jisook Moon, CHA University, Department of Biotechnology, Pangyo-Ro 335, Bundang-guSeongnam-siGyeonggi-do 463-400, Korea. Tel: 82-31-881-2027; Fax: 82-2-538-4102; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1159-1176, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688948
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved


Transcriptomic Analysis of Cultured Corneal Endothelial Cells as a Validation for Their Use in Cell Replacement Therapy

Ricardo F. Frausto, Derek J. Le, and Anthony J. Aldave

The Jules Stein Eye Institute, David Geffen School of Medicine at University of California, Los Angeles (UCLA), Los Angeles, CA, USA

The corneal endothelium plays a primary role in maintaining corneal homeostasis and clarity and must be surgically replaced with allogenic donor corneal endothelium in the event of visually significant dysfunction. However, a worldwide shortage of donor corneal tissue has led to a search for alternative sources of transplantable tissue. Cultured human corneal endothelial cells (HCEnC) have been shown to restore corneal clarity in experimental models of corneal endothelial dysfunction in animal models, but characterization of cultured HCEnC remains incomplete. To this end, we utilized next-generation RNA sequencing technology to compare the transcriptomic profile of ex vivo human corneal endothelial cells (evHCEnC) with that of primary HCEnC(pHCEnC) and HCEnC lines and to determine the utility of cultured and immortalized corneal endothelial cells as models of in vivo corneal endothelium. Multidimensional analyses of the transcriptome data sets demonstrated that primary HCEnC have a closer relationship to evHCEnC than do immortalized HCEnC. Subsequent analyses showed that the majority of the genes specifically expressed in HCEnC (not expressed in ex vivo corneal epithelium or fibroblasts) demonstrated a marked variability of expression in cultured cells compared with evHCEnC. In addition, genes associated with either corneal endothelial cell function or corneal endothelial dystrophies were investigated. Significant differences in gene expression and protein levels were observed in the cultured cells compared with evHCEnC for each of the genes tested except for AGBL1 and LOXHD1, which were not detected by RNA-seq or qPCR. Our transcriptomic analysis suggests that at a molecular level pHCEnC most closely resembleevHCEnC and thus represent the most viable cell culture-based therapeutic option for managing corneal endothelial cell dysfunction. Our findings also suggest that investigators should perform an assessment of the entire transcriptome of cultured HCEnCprior to determination of their potential clinical utility for the management of corneal endothelial cell failure.

Key words: Corneal endothelium; Transcriptome; Gene expression; Cell therapy; Human corneal endothelial cells (HCEnC)

Received February 19, 2015; final acceptance November 2, 2015. Online prepub date: September 2, 2015.
Address correspondence to Anthony J. Aldave, M.D., Professor of Ophthalmology, The Jules Stein Eye Institute, 100 Stein Plaza, UCLA, Los Angeles, CA 90095-7003, USA. Tel: +1-310-206-7202; Fax: +1-310-794-7906; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



Cell Transplantation, Vol. 25, pp. 1177-1191, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688957
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved


Facilitated Neural Differentiation of Adipose Tissue-Derived Stem Cells by Electrical Stimulation and Nurr-1 Gene Transduction

Yafeng Yang,*1 Teng Ma,*1 Jun Ge,*1 Xin Quan,* Le Yang,† Shu Zhu,* Liangliang Huang,* Zhongyang Liu,* Liang Liu,* Dan Geng,* Jinghui Huang,* and Zhuojing Luo*

*Institute of OrthopaedicsXijing Hospital, The Fourth Military Medical University, Xi’an, PR China
†Department of Pharmacology, School of Pharmacy, The Fourth Military Medical University, Xi’an, PR China

Neuron-like cells derived from adipose tissue-derived stem cells (ADSCs) have been considered one of the most promising cells for the treatment of neurodegenerative diseases and neurotrauma in the central nervous system (CNS). Thus far, extensive efforts have been made to facilitate neuronal differentiation of ADSCs, but limited progress has been achieved. In the present study, we tested the possibility of using a combination of electrical stimulation (ES) with Nurr-1 gene transduction to promote neuronal differentiation of ADSCs. The tolerance of ADSCs to ES was first examined by a cell apoptosis assay. The proliferation of cells was characterized using a CCK-8 assay. The morphology of cells was examined by scanning electron microscopy (SEM). The differentiation of ADSCs into neuron-like cells was examined by immunocytochemistry (ICC)–immunofluorescence staining, quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and enzyme linked immunosorbent assay (ELISA). The gene expression of microtubule-associated protein 2 (MAP-2), β-tubulin, neurofilament 200 (NF-200), octamer binding transcription factor 4 (OCT-4), and glial fibrillary acidic protein (GFAP) after stimulation was examined by qRT-PCR. We found that the optimal intensity of ES for neuronal differentiation of ADSCs was 1 V/cm. In addition, ES combined with Nurr-1 gene transduction increased the neuronal differentiation rate of ADSCs, the length of neurite-like processes, and the secretion of dopamine. Further studies showed that a combination of ES with Nurr-1 gene transduction was capable of promoting the expression of MAP-2, β-tubulin, and NF-200 but decreased the expression of OCT-4 and GFAP. All of these findings indicate that a combination of ES with Nurr-1 gene transduction could facilitate neuronal differentiation of ADSCs, which raises the possibility of its application in the treatment of neurodegenerative diseases and neurotrauma in the CNS.

Key words: Adipose tissue-derived stem cells (ADSCs); Electrical stimulation (ES); Nurr-1 gene; Neuronal differentiation; Polypyrrole/chitosan polymers; Lentivirus

Received November 4, 2014; final acceptance November 19, 2015. Online prepub date: September 2, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Jinghui Huang, Institute of OrthopaedicsXijing Hospital, The Fourth Military Medical University, No. 127 Changle West Road, Xi’an 710032, China. Tel: +86-29-84775285; Fax: +86-29-84775285; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Zhuojing Luo, Institute of OrthopaedicsXijing Hospital, The Fourth Military Medical University, No. 127 Changle West Road, Xi’an 710032, China. Tel: +86-29-84775285; Fax: +86-29-84775285; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1193-1206, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689442
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved


Comparative Efficacies of Long-Term Serial Transplantation of Syngeneic, Allogeneic, Xenogeneic, or CTLA4Ig-Overproducing Xenogeneic Adipose Tissue-Derived Mesenchymal Stem Cells on Murine Systemic Lupus Erythematosus

Eun Wha Choi,*† Hee Woo Lee,‡ Il Seob Shin,§ Ji Hyun Park,* Tae Won Yun,* Hwa Young Youn,‡ and Sung-Joo Kim*¶

*Laboratory Animal Research Center, Samsung Biomedical Research Institute, Gangnam-gu, Seoul, Republic of Korea
Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Republic of Korea
‡Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea
§Biostar Stem Cell Research Center, K-STEMCELL, Geumcheon-gu, Seoul, Republic of Korea
¶Department of Surgery, Division of Transplantation, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Republic of Korea

Allogeneic and xenogeneic transplantation are suitable alternatives for treating patients with stem cell defects and autoimmune diseases. The purpose of this study was to compare the effects of long-term serial transplantation of adipose tissue-derived mesenchymal stem cells (ASCs) from (NZB × NZW) F1 mice (syngeneic), BALB/c mice (allogeneic), or humans (xenogeneic) on systemic lupus erythematosus (SLE). The effects of transplanting human ASCs overproducing CTLA4Ig (CTLA4Ig-hASC) were also compared. Animals were divided into five experimental groups, according to the transplanted cell type. Approximately 500,000 ASCs were administered intravenously every 2 weeks from 6 to 60 weeks of age to all mice except for the control mice, which received saline. The human ASC groups (hASC and CTLA4Ig-hASC) showed a 13-week increase in average life spans and increased survival rates and decreased blood urea nitrogen, proteinuria, and glomerular IgG deposition. The allogeneic group also showed higher survival rates compared to those of the control, up to 40, 41, 42, 43, 44, 45, 52, and 53 weeks of age. Syngeneic ASC transplantation did not accelerate the mortality of the mice. The mean life span of both the syngeneic and allogeneic groups was prolonged for 6–7 weeks. Both human ASC groups displayed increased serum interleukin-10 and interleukin-4 levels, whereas both mouse ASC groups displayed significantly increased GM-CSF and interferon-γ levels in the serum. The strongest humoral immune response was induced by xenogeneic transplantation, followed by allogeneic, CTLA4Ig-xenogeneic, and syngeneic transplantations. Long-term serial transplantation of the ASCs from various sources displayed different patterns of cytokine expression and humoral responses, but all of them increased life spans in an SLE mouse model.

Key words: Systemic lupus erythematosus (SLE); Adipose tissue-derived mesenchymal stem cells; CTLA4Ig; (NZB × NZW) F1 mice

Received May 30, 2014; final acceptance November 9, 2015. Online prepub date: November 30, 2015.
Address correspondence to Eun Wha Choi, D. V. M., Ph.D., Laboratory Animal Research Center, Samsung Biomedical Research Institute, 81 Irwon-ro, Gangnam-gu, Seoul 135-710, Republic of Korea. Tel: +82-2-3410-3700; Fax: +82-2-3410-3628; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  orSung-Joo Kim, M.D., Ph.D., Professor, Department of Surgery, Division of Transplantation, Samsung Medical Center, Sungkyunkwan University School of Medicine, 81 Irwon-ro, Gangnam-gu, Seoul 135-710, Republic of Korea. Tel: +82-2-3410-3476; Fax: +82-2-3410-0040; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1207-1218, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690305
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved


Tolerogenic Dendritic Cells Generated by In Vitro Treatment With SAHA Are Not Stable In Vivo

Kristof Thewissen,*† Bieke Broux,*† Jerome J. A. Hendriks,*† Mandy Vanhees,‡ Piet Stinissen,*† Helena Slaets,*† and Niels Hellings*†

*Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium
†School of Life Sciences, Transnationale Universiteit Limburg, Diepenbeek, Belgium
‡Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium

The aim of this study is to examine whether the histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), can generate dendritic cells (DCs) with a stable tolerogenic phenotype to counteract autoimmune responses in an animal model of multiple sclerosis. We investigated if the tolerogenic potency of DCs could be increased by continuous treatment during in vitro differentiation toward DCs compared to standard 24-h in vitro treatment of already terminally differentiated DCs. We show that in vitro treatment with SAHA reduces the generation of new CD11c+
DCs out of mouse bone marrow. SAHA-generated DCs show reduced antigen-presenting function as evidenced by a reduction in myelin endocytosis, a decreased MHC II expression, and a failure to upregulate costimulatory molecules upon LPS challenge. In addition, SAHA-generated DCs display a reduction in proinflammatory cytokines and molecules involved in apoptosis induction, inflammatory migration, and TLR signaling, and they are lessimmunostimulatory compared to untreated DCs. We demonstrated that the underlying mechanism involves a diminished STAT1 phosphorylation and was independent of STAT6 activation. Although in vitro results were promising, SAHA-generated DCs were not able to alleviate the development of experimental autoimmune encephalomyelitis in mice. In vitro washout experiments demonstrated that the tolerogenic phenotype of SAHA-treated DCs is reversible. Taken together, while SAHA potently booststolerogenic properties in DCs during the differentiation process in vitro, SAHA-generated DCs were unable to reduce autoimmunity in vivo. Our results imply that caution needs to be taken when developing DC-based therapies to induce tolerance in the context of autoimmune disease.

Key words: Dendritic cells (DCs); Multiple sclerosis; Histone deacetylase inhibitor; Anti-inflammatory; Tolerance

Received June 16, 2014; final acceptance February 9, 2016. Online prepub date: December 18, 2015.
Address correspondence to Niels Hellings, Biomedical Research Institute, Hasselt University, Martelarenlaan 42, B3500 Hasselt, Belgium. Tel: +32-11-269268; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it