Cell Transplantation 19(10) Abstracts

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Cell Transplantation, Vol. 19, pp. 1217–1223, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X507187
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Review

Adipocyte Transplantation and Stem Cells: Plastic Surgery Meets Regenerative Medicine

Carlo Tremolada,* Giancarlo Palmieri,† and Camillo Ricordi‡

*Istituto Image and San Paolo Hospital, Milan, Italy
†Niguarda Hospital, Milan, Italy
‡Cell Transplant Center, Diabetes Research Institute, University of Miami, Miami, FL, USA

The technologies for adipose tissue harvesting, processing, and transplantation have substantially evolved in the past two decades. Clinically driven advancements have paralleled a significant improvement in the understanding of cellular, molecular, and immunobiological events surrounding cell and tissue transplantation. These new mechanistic insights could be of assistance to better understand the mechanisms underlying some of the observed clinical improvements. In addition to plastic and reconstructive surgical applications, adipose tissue has become central to an increasing number of translational efforts involving adipose tissue-derived progenitor cells. The growing interest in this area of research has resulted in the exploration of many novel research and clinical applications that utilize adipose tissue grafting and/or progenitor/stem cell-derived cell products obtained from this tissue source. Progenitor, endothelial, and mesenchymal stem cells derived from adipose tissue could therefore not only be central to plastic and reconstructive surgery applications, but also become the focus of an array of therapeutic solutions for many disease conditions, such as those affecting bone, cartilage, muscle, liver, kidney, cardiac, neural, and the pancreas, expanding the possible indications and translational potential of tissue, cell-based, and regenerative medicine strategies.

Key words: Stem cell; Adipose; Fat; Graft; Tissue remodeling; Lipofilling

Address correspondence to Camillo Ricordi, M.D., Cell Transplant Center and Diabetes Research Institute, University of Miami, 1450 N. W. 10th Avenue, Miami, FL 33136, USA. Tel: (305) 243-6913; Fax: (305) 243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1225–1235, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/09638910X519265
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Purification and Characterization of Adipose-Derived Stem Cells From Patients With Lipoaspirate Transplant

Valentina Folgiero,*1 Emilia Migliano,†1 Marinella Tedesco,† Stefano Iacovelli,‡ Giulia Bon,* Maria Luisa Torre,§ Ada Sacchi,* Mario Marazzi,¶ Stefania Bucher,† and Rita Falcioni*

*Dipartimento di Oncologia Sperimentale, Istituto Regina Elena, Roma, Italy
†Divisione di Chirurgia Plastica e Ricostruttiva, Istituto San Gallicano, Rome, Italy
‡Dipartimento di Biotecnologie Cellulari ed Ematologia, Università di Roma “La Sapienza”, Roma, Italy
§Dipartimento di Chimica Farmaceutica, Università di Pavia, Pavia, Italy
¶Unità di Terapia Tissutale, Azienda Ospedaliera Niguarda Ca’ Granda, Milano, Italy

Techniques for medical tissue regeneration require an abundant source of human adult stem cells. There is increasing evidence that adipose stem cells contribute to restoration of tissue vascularization and organ function. The object of our study was to isolate and characterize adult adipose-derived stem cells from patients undergoing on lipoaspirate transplant with the aim to improve tissue regeneration. Adipose-derived stem cells were isolated and purified from the lipoaspirate of 15 patients and characterized for CD markers and the ability to differentiate toward the adipogenic lineage. We found that purified adipose stem cells express high level of CD49d, CD44, CD90, CD105, CD13, and CD71 and these markers of staminality were maintained at high level for at least 3 months and seven passages of in vitro culture. As expected, these cells resulted negative for the endothelial and hematopoietic-specific markers CD31, CD106, CD34, and CD45. Differentiation towards adipogenic lineage demonstrated that purified adipose-derived stem cells are still able to become adipocytes at least 3 months after in vitro culture. The analysis of Akt and MAPK phosphorylation confirmed a modulation of their activity during differentiation. Interestingly, we established for the first time that, among the p53 family members, a strong upregulation of p63 expression occurs in adipocytic differentiation, indicating a role for this transcription factor in adipocytic differentiation. Taken together, these data indicate that purified lipoaspirate-derived stem cells maintain their characteristic of staminality for a long period of in vitro culture, suggesting that they could be applied for cell-based therapy to improve autologous lipoaspirate transplant.

Key words: Lipoaspirate transplant; Tissue regeneration; Stem cells; Adipocyte differentiation; p53 family

1
These authors provided equal contribution.
Address correspondence to Rita Falcioni, Department of Experimental Oncology, Molecular Oncogenesis Laboratory, Regina Elena Cancer Institute, Via delle Messi d’Oro, 156, 00158 Rome, Italy. Tel: +39 06 52662535; Fax: +39 06 52662505; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Stefania Bucher, Division of Plastic and Reconstructive Surgery, San Gallicano Institute, Via Elio Chianesi, 54, 00144 Rome, Italy. Tel: +39 06 52665901; Fax: +39 06 52664858; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1237–1246, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X506863
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Autologous Bone Marrow Infusion Activates the Progenitor Cell Compartment in Patients With Advanced Liver Cirrhosis

Ja Kyung Kim,*,**††1 Young Nyun Park,#**††1 Jin Seok Kim,* Mi-Suk Park,‡** Yong Han Paik,*#**†† Jae-Yeon Seok,† Yong Eun Chung,‡ Hyun Ok Kim,§ Kyung Sik Kim,¶** Sang Hoon Ahn,*,**†† Do Young Kim,*,**†† Myeong-Jin Kim,‡** Kwan Sik Lee,*,**†† Chae Yoon Chon,*,**†† Soo Jeong Kim,* Shuji Terai,‡‡ Isao Sakaida,‡‡ and Kwang-Hyub Han*#**††

*Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea
†Department of Pathology, Yonsei University College of Medicine, Seoul, Korea
‡Department of Radiology, Yonsei University College of Medicine, Seoul, Korea
§Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea
¶Department of Surgery, Yonsei University College of Medicine, Seoul, Korea
#BK 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
**Yonsei Institute of Gastroenterology, Yonsei University College of Medicine, Seoul, Korea
††Liver Cirrhosis Clinical Research Center, Seoul, Korea
‡‡Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan

Several clinical trials of bone marrow cell infusion in patients with liver cirrhosis (LC) have shown clinical improvement, despite conflicting results from animal models. We investigated serial pathological features and the clinical impact after autologous bone marrow infusion (ABMI) in patients with advanced LC. Ten patients with advanced LC due to chronic hepatitis B virus infection underwent ABMI. Serological tests, MRI, and liver biopsies were performed, and quality of life was assessed by a questionnaire. Median serum albumin and hemoglobin levels increased significantly after ABMI. All patients showed an improvement in quality of life, with no serious adverse events. Liver volume, measured by MRI, increased in 80% of the patients, and ascites decreased after ABMI. Child-Pugh scores were also significantly improved at 6 months after ABMI. In the serially biopsied livers, a gradually increasing activation of the hepatic progenitor cell (HPC) compartment, including HPC activation (ductular reaction) and HPC differentiation (intermediate hepatocyte), reached a peak after 3 months, with continued proliferation of hepatocytes, and returned to baseline levels after 6 months. There was no significant change in grade or stage of liver fibrosis or stellate cell activation after ABMI. ABMI is suggested to improve liver function and to activate the progenitor cell compartment. Although clinical improvement was sustained for more than 6 months, histological changes in the liver returned to baseline 6 months after ABMI. Further comparative studies are warranted.

Key words: Adult bone marrow stem cell; Autologous bone marrow transplantation; Liver cirrhosis; Liver regeneration; Progenitor cell

1
These authors provided equal contribution to this study.
Address correspondence to Kwang-Hyub Han, M.D., Department of Internal Medicine, Yonsei University College of Medicine, 250 Seongsanno Seodaemun-gu, Seoul, 120-752, Korea. Tel: 82-2-2228-1949; Fax: 82-2-393-6884; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1247–1260, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X505864
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Human Bone Marrow-Derived CD133
+ Cells Delivered to a Collagen Patch on Cryoinjured Rat Heart Promote Angiogenesis and Arteriogenesis

M. Pozzobon,* S. Bollini,* L. Iop,† P. De Gaspari,* A. Chiavegato,† C. A. Rossi,* S. Giuliani,‡ F. Fascetti Leon,‡ N. Elvassore,§ S. Sartore,¶ and P. De Coppi‡#

*Stem Cell Processing Laboratory, Cord Blood Bank, Department of Pediatric Oncohematology and Stem Cell Unit, University of Padua, Padua, Italy
†Department of Biomedical Sciences, University of Padua, Padua, Italy
‡Department of Pediatric Surgery, University of Padua, Padua, Italy
§Department of Chemical Engineering, University of Padua, Padua, Italy
¶Stem Cell Unit, University of Padua, Padua, Italy
#Surgery Unit, UCL Institute of Child Health and Great Ormond Street Hospital, London, UK

Transplanting hematopoietic and peripheral blood-derived stem/progenitor cells can have beneficial effects in slowing the effects of heart failure. We investigated whether human bone marrow CD133+-derived cells (BM-CD133+ cells) might be used for cell therapy of heart injury in combination with tissue engineering. We examined these cells for: 1) their in vitro capacity to be converted into cardiomyocytes (CMs), and 2) their potential for in vivo differentiation when delivered to a tissue-engineered type I collagen patch placed on injured hearts (group II). To ensure a microvascular network ready for use by the transplanted cells, cardiac injury and patching were scheduled 2 weeks before cell injection. The cardiovascular potential of the BM-CD133+ cells was compared with that of a direct injection (group I) of the same cells in heart tissue damaged according to the same schedule as for group II. While a small fraction (2 ± 0.5%) of BMCD133+ cells cocultured with rat CMs switched in vitro to a CM-like cell phenotype, in vivo—and in both groups of nude rats transplanted with BM-CD133+—there was no evidence of any CM differentiation (as detected by cardiac troponin I expression), but there were signs instead of new capillaries and small arterioles. While capillaries prevailed over arterioles in group II, the opposite occurred in group I. The transplanted cells further contributed to the formation of new microvessels induced by the patch (group II) but the number of vessels did not appear superior to the one developed after directly injecting the BM-CD133+ cells into the injured heart. Although chimeric human–rat microvessels were consistently found in the hearts of both groups I and II, they represented a minority (1.5–2.3%) compared with those of rat origin. Smooth muscle myosin isoform expression suggested that the arterioles achieved complete differentiation irrespective of the presence or absence of the collagen patch. These findings suggest that: 1) BM-CD133+ cells display a limited propensity for in vitro conversion to CMs; 2) the preliminarily vascularized bioscaffold did not confer a selective homing and differentiation advantage for the phenotypic conversion of BM-CD133+ cells into CMs; and 3) combined patching and cell transplantation is suitable for angiogenesis and arteriogenesis, but it does not produce better results, in terms of endothelial and smooth muscle cell differentiation, than the “traditional” method of cell injection into the myocardium.

Key words: Stem cells; Xenotransplantation; Collagen; Cardiac tissue engineering

Address correspondence to Michela Pozzobon, Stem Cell Processing Laboratory, Department of Pediatric Oncohematology, University of Padova, Via Orus 2, 35129 Padua, Italy. Tel: +39 049 7923230; Fax: +39 049 7923250; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1261–1279, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X507204
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Nonsenescent Hsp27-Upregulated MSCs Implantation Promotes Neuroplasticity in Stroke Model

Shih-Ping Liu,*† Dah-Ching Ding,‡ Hsiao-Jung Wang,* Ching-Yuan Su,§ Shinn-Zong Lin,*¶ Hung Li,§1 and Woei-Cherng Shyu*¶

*Center for Neuropsychiatry, China Medical University Hospital, Taichung, Taiwan
†Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan
‡Department of Obstetrics and Gynecology, Buddhist Tzu-Chi General Hospital, Hualien, Taiwan
§Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
¶Graduate Institute of Immunology, China Medical University, Taichung, Taiwan
#China Medical University Beigang Hospital, Yunlin, Taiwan

Cellular senescence induces changes in cellular physiology, morphology, proliferative capacity, and gene expression. Stem cell senescence might be one of the major issues of limited efficacy of stem cell transplantation. In this study, we demonstrated that implantation of human umbilical cord mesenchymal stem cells (hUCMSCs) cultured in human umbilical cord serum (hUCS) significantly enhanced neuroplasticity and angiogenesis in stroke and ischemic limb models. Immunophenotypic analysis indicated that hUCMSCs cultured in hUCS had more small and rapidly self-renewing cells than those expanded in FCS. The main cause of greater senescence in FCS-cultured cells was increased generation of reactive oxygen species (ROS). Proteome profiling showed significantly more senescence-associated vimentin in FCS-cultured hUCMSCs than in hUCS-cultured hUCMSCs. In contrast, there was significant upregulation of heat shock protein 27 (Hsp27) in the hUCS-cultured hUCMSCs. By gene targeting, we found that overexpression of Hsp27 may downregulate vimentin expression through inhibition of the nuclear translocation of p65 (NF-κB signaling). Thus, an interaction between Hsp27 and vimentin may modulate the degree of senescence in hUCS- and FCS-cultured hUCMSCs. In summary, hUCMSCs exhibiting senescence are detrimental to cell engraftment and differentiation in animal models via activation of NF-κB pathway. Human stem cells incubated in hUCS might reduce the senescent process through upregulation of Hsp27 to increase implantation efficiency.

Key words: Human umbilical cord mesenchymal stem cells (UCMSCs); Human umbilical cord serum (hUCS); Fetal calf serum (FCS); Senescence; Vimentin; Heat shock protein 27 (Hsp27); Reactive oxygen species (ROS); NF-κB; Stroke; Ischemic limbs

1
Hung Li recently passed away. (This work was done primarily in Academia Sinica and supported by grant award to Dr. Hung Li)
Address correspondence to Woei-Cherng Shyu, MD., Ph.D., Center for Neuropsychiatry, China Medical University and Hospital, Taichung, Taiwan. Tel: 886-4-22052121, ext. 7811; Fax: 886-4-22052121, ext. 7810; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Shinn-Zong Lin, M.D., Ph.D., Center for Neuropsychiatry, China Medical University and Hospital, Taichung, Taiwan. Tel: 886-4-22052121, ext. 6034; Fax: 886-4-220806666; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1281–1290, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X509040
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Treatment of Ocular Surface Injuries by Limbal and Mesenchymal Stem Cells Growing on Nanofiber Scaffolds

Alena Zajicova,* Katerina Pokorna,*† Anna Lencova,* Magdalena Krulova,*† Eliska Svobodova,*† Sarka Kubinova,‡ Eva Sykova,‡ Martin Pradny,§ Jiri Michalek,§ Jana Svobodova,¶ Marcela Munzarova,¶ and Vladimir Holan*†

*Institute of Molecular Genetics, Academy of Sciences, Prague, Czech Republic
†Faculty of Science, Charles University, Prague, Czech Republic
‡Institute of Experimental Medicine, Academy of Sciences, Prague, Czech Republic
§Institute of Macromolecular Chemistry, Academy of Sciences, Prague, Czech Republic
¶Elmarco, Liberec, Czech Republic

Stem cell (SC) therapy represents a promising approach to treat a wide variety of injuries, inherited diseases, or acquired SC deficiencies. One of the major problems associated with SC therapy remains the absence of a suitable matrix for SC growth and transfer. We describe here the growth and metabolic characteristics of mouse limbal stem cells (LSCs) and mesenchymal stem cells (MSCs) growing on 3D nanofiber scaffolds fabricated from polyamide 6/12 (PA6/12). The nanofibers were prepared by the original needleless electrospun Nanospider technology, which enables to create nanofibers of defined diameter, porosity, and a basis weight. Copolymer PA6/12 was selected on the basis of the stability of its nanofibers in aqueous solutions, its biocompatibility, and its superior properties as a matrix for the growth of LSCs, MSCs, and corneal epithelial and endothelial cell lines. The morphology, growth properties, and viability of cells grown on PA6/12 nanofibers were comparable with those grown on plastic. LSCs labeled with the fluorescent dye PKH26 and grown on PA6/12 nanofibers were transferred onto the damaged ocular surface, where their seeding and survival were monitored. Cotransfer of LSCs with MSCs, which have immunosuppressive properties, significantly inhibited local inflammatory reactions and supported the healing process. The results thus show that nanofibers prepared from copolymer PA6/12 represent a convenient scaffold for growth of LSCs and MSCs and transfer to treat SC deficiencies and various ocular surface injuries.

Key words: Limbal stem cells (LSCs); Mesenchymal stem cells (MSCs); Nanofiber scaffolds; Ocular surface injuries; Inflammation; Tissue regeneration

Address correspondence to Vladimir Holan, Ph.D., Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague 4, Czech Republic. Tel: +420-241063226; Fax: +420-224310955; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1291–1306, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X505477
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Immortalized Human Fetal Retinal Cells Retain Progenitor Characteristics and Represent a Potential Source for the Treatment of Retinal Degenerative Disease

Shazeen M. Hasan,*1 Anthony A. Vugler,*1 Erik A. Miljan,† John D. Sinden,† Stephen E. Moss,* and John Greenwood*

*Department of Cell Biology, UCL Institute of Ophthalmology, London, UK
†ReNeuron, Guildford, UK

Human fetal retinal cells have been widely advocated for the development of cellular replacement therapies in patients with retinal dystrophies and age-related macular degeneration. A major limitation, however, is the lack of an abundant and renewable source of cells to meet therapeutic demand, although theoretically this may be addressed through the use of immortalized retinal progenitor cell lines. Here, we have used the temperature-sensitive tsA58 simian virus SV40 T antigen to conditionally immortalize human retinal progenitor cells isolated from retinal tissue at 10–12 weeks of gestation. We show that immortalized human fetal retinal cells retain their progenitor cell properties over many passages, and are comparable with nonimmortalized human fetal retinal cultures from the same gestational period with regard to expression of certain retinal genes. To evaluate the capacity of these cells to integrate into the diseased retina and to screen for potential tumorigenicity, cells were grafted into neonatal hooded Lister rats and RCS dystrophic rats. Both cell lines exhibited scarce integration into the host retina and failed to express markers of mature differentiated retinal cells. Moreover, although immortalized cells showed a greater propensity to survive, the cell lines demonstrated poor long-term survival. All grafts were infiltrated with host macrophage/microglial cells throughout their duration of survival. This study demonstrates that immortalized human fetal retinal progenitor cells retain their progenitor characteristics and may therefore have therapeutic potential in strategies that demand a renewable and consistent supply of donor cells for the treatment of degenerative retinal diseases.

Key words: Retina; Progenitor cells; Conditional immortalization; Cellular therapy

1
These authors provided equal contribution to this study.
Address correspondence to John Greenwood, Department of Cell Biology, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK. Tel: +44 (0)207 6086858; Fax: +44 (0)207 6086810; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1307–1323, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X509059
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Glutamatergic Excitation and GABA Release From a Transplantable Cell Line

Jana Mejía-Toiber,* José Abraham Márquez-Ramos,† Mauricio Díaz-Muñoz,‡ Fernando Peña,† Manuel B. Aguilar,‡ and Magda Giordano*

*Departamento de Neurobiología Conductual y Cognitiva, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, México
†Departamento de Farmacobiología, CINVESTAV-Sur. Calzada de los Tenorios 235, Delegación Tlalpan, México
‡Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, México

"The cell line M213-2O CL-4 was derived from cell line M123-2O and further modified to express human glutamate decarboxylase (hGAD-67), the enzyme that synthesizes GABA. Brain transplants of this cell line in animal models of epilepsy have been shown to modulate seizures. However, the mechanisms that underlie such actions are unknown. The purpose of the present study was to characterize this cell line and its responsiveness to several depolarizing conditions, in order to better understand how these cells exert their effects. Intracellular GABA levels were 34-fold higher and GAD activity was 16-fold higher in clone M213-2O CL-4 than in M123-2O. Both cell lines could take up [3H]GABA in vitro, and this uptake was prevented by nipecotic acid. By combining GABA release measurements and calcium imaging in vitro, we found that high extracellular K+, zero Mg2+, or glutamate activated M213-2O CL-4 cells and resulted in GABA release. The response to glutamate appeared to be mediated by AMPA/NMDA-like receptors. High KCl-induced GABA release was prevented when a Ca2+-free Krebs solution was used, suggesting an exocytotic-like mechanism. These results indicate that the cell line M213-2O CL-4 synthesizes, releases, and takes up GABA in vitro, and can be activated by depolarizing stimuli.

Key words: Glutamate decarboxylase (GAD); Cell therapy; GABA uptake; Exocytosis; Calcium transients

Address correspondence to Magda Giordano, Departamento de Neurobiología Conductual y Cognitiva, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, Qro. 76230, México. Tel: +52-442-238-1061; Fax: +52-442-238-1046; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1325–1337, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X505855
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Synergistic Effect of Neural Stem Cells and Olfactory Ensheathing Cells on Repair of Adult Rat Spinal Cord Injury

Gan Wang,*‡ Qiang Ao,† Kai Gong,* Huancong Zuo,† Yandao Gong,* and Xiufang Zhang*

*State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Life Sciences, Tsinghua University, Beijing, China
†Institute of Neurological Disorders, Yuquan Hospital, Tsinghua University, Beijing, China
‡Department of Chemistry and Biology, College of Science, National University of Defense Technology, Changsha, China

Spinal cord injury (SCI) is a common clinical disease that places a heavy burden on families and society. Cellular therapy provides a method of giving a supplement of cells lost in the injury and promoting functional recovery after SCI. Neural stem cells (NSCs) and olfactory ensheathing cells (OECs) are two most promising cell types. NSCs have the potential of differentiating into neurons and glial cells, and OECs could help the axons of neurons pass through the glial scar to promote functional recovery. NSCs were isolated from the cortices of fetal rats on days 12–14 of embryonic development and OECs were isolated from the olfactory bulbs of adult rats. In vitro coculture studies demonstrated OECs could promote NSCs to differentiate into neurons. Four groups of rats that had been 3/4 spinal cord transectioned at T9 were injected with DMEM/F12 solution, NSCs, OECs, and NSCs + OECs, respectively, 7 days post-SCI. Twelve weeks postoperation, the hindlimb locomotor function of rats in the cotransplantation group was significantly improved compared with that in the other three groups. Histological observation and immunohistochemical staining of NF-200 both showed new nerve fibers across the injured region. Cotransplantation of NSCs and OECs might have a synergistic effect on promoting neural regeneration and improving the recovery of locomotion function. Cotransplantation of NSCs and OECs was better than a single graft of either NSCs or OECs. These findings have provided a new way of thinking in the treatment of SCI.

Key words: Spinal cord injury; Neural stem cells; Olfactory ensheathing cells; Cell transplantation

Address correspondence to Xiufang Zhang, School of Life Sciences, Tsinghua University, Beijing 100084, China. Tel: +86-10-62783261; Fax: +86-10-62794214; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Qiang Ao, Institute of Neurological Disorders, Yuquan Hospital, Tsinghua University, Beijing 100049, China. Tel: +86-10-88255960; Fax: +86-10-62794214; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1339–1348, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X507196
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Use of a Collagen–Elastin Matrix as Transport Carrier System to Transfer Proliferating Epidermal Cells to Human Dermis In Vitro

Taco Waaijman,* Melanie Breetveld,* Magda Ulrich,†‡ Esther Middelkoop,†‡ Rik J. Scheper,§ and Susan Gibbs*

*Department of Dermatology, VU University Medical Centre, Amsterdam, The Netherlands
†Plastic, Reconstructive & Hand Surgery, VU University Medical Centre, Amsterdam, The Netherlands
‡Association of Dutch Burn Centers, Beverwijk, The Netherlands
§Department of Pathology, VU University Medical Centre, Amsterdam, The Netherlands

This in vitro study describes a novel cell culture, transport, and transfer protocol that may be highly suitable for delivering cultured proliferating keratinocytes and melanocytes to large open skin wounds (e.g., burns). We have taken into account previous limitations identified using other keratinocyte transfer techniques, such as regulatory issues, stability of keratinocytes during transport (single cell suspensions undergo terminal differentiation), ease of handling during application, and the degree of epidermal blistering resulting after transplantation (both related to transplanting keratinocyte sheets). Large numbers of proliferating epidermal cells (EC) (keratinocytes and melanocytes) were generated within 10–14 days and seeded onto a threedimensional matrix composed of elastin and collagen types I, III, and V (Matriderm®), which enabled easy and stable transport of the EC for up to 24 h under ambient conditions. All culture conditions were in accordance with the regulations set by the Dutch Central Committee on Research Involving Human Subjects (CCMO). As an in vitro model system for clinical in vivo transfer, the EC were then transferred from Matriderm onto human acellular dermis during a period of 3 days. After transfer the EC maintained the ability to regenerate into a fully differentiated epidermis containing melanocytes on the human dermis. Proliferating keratinocytes were located in the basal layer and keratin-10 expression was located in differentiating suprabasal layers similar to that found in human epidermis. No blistering was observed (separation of the epidermis from the basement membrane). Keratin-6 expression was strongly upregulated in the regenerating epidermis similar to normal wound healing. In summary, we show that EC-Matriderm contains viable, metabolically active keratinocytes and melanocytes cultured in a manner that permits easy transportation and contains epidermal cells with the potential to form a pigmented reconstructed epidermis. This in vitro study has produced a robust protocol that is ready for clinical studies in the future.

Key words: Transfer; Keratinocyte; Melanocyte; Burns; Wound healing

Address correspondence to Susan Gibbs, Department of Dermatology, VU University Medical Centre, De Boelelaan, 1081 HV Amsterdam, The Netherlands. Tel: 0031 20 4442815; Fax: 0031 20 4442816; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1349–1357, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X506854
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Plasticity of Fetal Cartilaginous Cells

Aurelie Quintin,*† Constantin Schizas,‡ Corinne Scaletta,*‡ Sandra Jaccoud,*† Lee Ann Applegate,*‡ and Dominique P. Pioletti†

*Cellular Therapy Unit, Department of Musculoskeletal Medicine, University Hospital Center and University of Lausanne, CHUV-UNIL, Lausanne, Switzerland
†Laboratory of Biomechanical Orthopedics, Institute of Bioengineering, Ecole Polytechnique Fe´de´rale de Lausanne (EPFL), Lausanne, Switzerland
‡Department of Musculoskeletal Medicine, University Hospital Center and University of Lausanne, CHUV-UNIL, Lausanne, Switzerland

Tissue-specific stem cells found in adult tissues can participate in the repair process following injury. However, adult tissues, such as articular cartilage and intervertebral disc, have low regeneration capacity, whereas fetal tissues, such as articular cartilage, show high regeneration ability. The presence of fetal stem cells in fetal cartilaginous tissues and their involvement in the regeneration of fetal cartilage is unknown. The aim of the study was to assess the chondrogenic differentiation and the plasticity of fetal cartilaginous cells. We compared the TGF-β3-induced chondrogenic differentiation of human fetal cells isolated from spine and cartilage tissues to that of human bone marrow stromal cells (BMSC). Stem cell surface markers and adipogenic and osteogenic plasticity of the two fetal cell types were also assessed. TGF-β3 stimulation of fetal cells cultured in high cell density led to the production of aggrecan, type I and II collagens, and variable levels of type X collagen. Although fetal cells showed the same pattern of surface stem cell markers as BMSCs, both type of fetal cells had lower adipogenic and osteogenic differentiation capacity than BMSCs. Fetal cells from femoral head showed higher adipogenic differentiation than fetal cells from spine. These results show that fetal cells are already differentiated cells and may be a good compromise between stem cells and adult tissue cells for a cell-based therapy.

Key words: Stem cells; Fetal cells; Chondrogenesis; Plasticity

Address correspondence to Prof. Dominique P. Pioletti, Laboratory of Biomechanical Orthopedics, Station 15, 1015 Lausanne, Switzerland. Tel: 41 21 693 8341; Fax: 41 21 693 8660; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it