Cell Medicine 7(1) Abstracts

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Cell Medicine, Vol. 7, pp. 1–13, 2014
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DOI: http://dx.doi.org/10.3727/215517914X680056
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2014 Cognizant Comm. Corp.
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Review

Application of Induced Pluripotent Stem Cells in Liver Diseases

Yue Yu,*† Xuehao Wang,*† and Scott L. Nyberg‡

*Key Laboratory of Living Donor Liver Transplantation, Ministry of Public Health, China
†Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
‡Division of Experimental Surgery, Mayo Clinic College of Medicine, Rochester, MN, USA

Tens of millions of patients are affected by liver disease worldwide. Many of these patients can benefit from therapy involving hepatocyte transplantation. Liver transplantation is presently the only proven treatment for many medically refractory liver diseases including end-stage liver failure and inherited metabolic liver disease. However, the shortage in transplantable livers prevents over 40% of listed patients per year from receiving a liver transplant; many of these patients die before receiving an organ offer or become too sick to transplant. Therefore, new therapies are needed to supplement whole-organ liver transplantation and reduce mortality on waiting lists worldwide. Furthermore, the remarkable regenerative capacity of hepatocytes in vivo is exemplified by the increasing number of innovative cell-based therapies and animal models of human liver disorders. Induced pluripotent stem cells (iPSCs) have similar properties to those of embryonic stem cells (ESCs) but bypass the ethical concerns of embryo destruction. Therefore, generation of hepatocyte-like cells (HLCs) using iPSC technology may be beneficial for the treatment of severe liver diseases, screening of drug toxicities, basic research of several hepatocytic disorders, and liver transplantation. Here we briefly summarize the growing number of potential applications of iPSCs for treatment of liver disease.

Key words: Induced pluripotent stem cells (iPSCs); Liver diseases; Cell therapy; Hepatocyte transplantation

Received November 12, 2013; final acceptance April 7, 2014. Online prepub date: April 22, 2014.
Address correspondence to Scott L. Nyberg, M.D., Ph.D., Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. Tel: +1-507-266-6772; Fax: +1-507-266-2810; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Yue Yu, M.D., Ph.D., The First Affiliated Hospital, Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu Province 210029, China. Tel: +86-25-68136212; Fax: +86-25-8367 2106; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 7, pp. 15–24, 2014
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DOI: http://dx.doi.org/10.3727/215517914X681785
Copyright ©
2014 Cognizant Comm. Corp.
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Integration-Free Human Induced Pluripotent Stem Cells From Type 1 Diabetes Patient Skin Fibroblasts Show Increased Abundance of Pancreas-SpecificmicroRNAs

Jun Liu,*1 Mugdha V. Joglekar,† Huseyin Sumer,* Anandwardhan A. Hardikar,† Halena Teede,‡§ and Paul J. Verma

*Cell Reprogramming and Stem Cells, Monash Institute of Medical Research, Monash University, Clayton, VIC, Australia
†Diabetes and Islet Biology Group, NHMRC Clinical Trials Centre, The University of Sydney, Camperdown, NSW, Australia
‡School of Public Health and Preventive Medicine, Monash University, Clayton, VIC, Australia
§Diabetes Unit, Southern Health Clayton, Clayton, VIC, Australia
Turretfield Research Centre, South Australian Research and Development Institute, Rosedale SA 3050, Australia

Type 1 diabetes (T1D) is a disease that is typically associated with multigenetic changes as well as environmental triggers. Disease-specific induced pluripotent stem cells (iPSCs) are preferable cell sources to study T1D, as they are derived from patient cells and therefore capture the disease genotype in a stem cell line. The purpose of this study was to generate integration-free iPSCs from adult skin fibroblasts with T1D. iPSCs were generated by transfection of synthetic mRNAs encoding transcription factors OCT4, SOX2, KLF4, c-MYC, and LIN28. Phase-contrast microscopy, immunocytochemistrykaryotypingbisulfite genomic sequencing, reverse transcription-polymerase chain reaction, and teratoma formation assay were used to determine reprogramming efficiency, pluripotency, and differentiation potential. Following 18 consecutive days of synthetic mRNA transfections, the T1D patient skin fibroblasts underwent morphological changes, and the aggregated clumps exhibited a human embryonic stem cell (ESC)-like morphology with a high nucleus/cytoplasm ratio. Highly efficient generation of iPSCs was achieved using the mRNA reprogramming approach. The disease-specific iPSCs expressed pluripotency markers, maintained a normal karyotype, and formed teratomas containing tissues representative of the three germ layers when injected into immune-deficient mice. Of interest, the iPSCs showed upregulations of pancreas-specific microRNAs, compared with parental fibroblasts. These data indicate that T1D patient skin fibroblasts can be reprogrammed to pluripotency using a synthetic mRNA approach. These cells can serve as a useful tool for the identification of genes that are involved in autoimmune reactions as well as generating patient-matched b-cells for cell-based therapy.

Key words: Type 1 diabetes (T1D); Induced pluripotent stem cells (iPSCs); Pluripotency reprogramming; microRNA

Received February 20, 2014; final acceptance April 29, 2014. Online prepub date: May 2, 2014.
1Current address: Department of Biological Engineering, Monash University, Clayton, VIC, Australia
Address correspondence to Prof. Paul VermaTurretfield Research Centre, South Australian Research and Development Institute, Rosedale SA 3050, Australia. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr. Jun Liu, Cell Reprogramming and Stem Cells, Monash Institute of Medical Research, Monash University, Clayton VIC 3168, Australia. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 7, pp. 25–35, 2014
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DOI: http://dx.doi.org/10.3727/215517914X679265
Copyright ©
2014 Cognizant Comm. Corp.
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Human Menstrual Blood-Derived Mesenchymal Cells as New Human Feeder Layer System for Human Embryonic Stem Cells

Danubia Silva dos Santos, Vanessa Carvalho Coelho de Oliveira, Karina Dutra Asensi, Leandro Vairo, Adriana Bastos Carvalho, Antonio Carlos Campos de Carvalho, and Regina Coeli dos Santos Goldenberg

Instituto de Biofisica Carlos Chagas FilhoUniversidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

Human embryonic stem cells (hESCs) in general require coculture with feeder layers in order to remain undifferentiated. However, the use of animal-derived feeder layers is incompatible with the clinical setting. The objective of this work was to investigate whether human menstrual blood-derived mesenchymal cells (MBMCs) can substitute mouse embryonic fibroblasts (MEFs) as a feeder layer for H9-hESCs. Both feeder cell types were isolated and cultured in DMEM F-12 and high glucose DMEM, respectively. After three passages, they were inactivated with mitomycin C. To test MBMC feeder layer capacity, hESCs were grown over MBMCs and MEFs under standard conditions. hESC growth, proliferation, survival, and maintenance of the undifferentiated state were evaluated. hESCs grown over MBMCs preserved their undifferentiated state presenting standard morphology, expressing alkaline phosphatase, transcription factors OCT3/4SOX2, and NANOG by RT-PCR and SSEA-4 and OCT3/4 by immunofluorescence assays. It is noteworthy that none of the feeder cells expressed these proteins. The average colony size of the hESCs on MBMCs was higher when compared to MEFs (p < 0.05; mean ± SD, n = 3). Growth factor analysis revealed amplification of the transcripts for FGF-2BMP4TGF-b, VEGF, and PEDF by RT-PCR in MBMCs and MEFs before and after inactivation. Furthermore, similar embryoid body formation, size, and morphology were observed in both feeder layers. In addition, EBs expressed marker genes for the three germ layers cultured on both feeder cells. In conclusion, MBMCs are able to maintain hESCs in an undifferentiated state with comparable efficiency to MEFs. Therefore, MBMCs are a suitable alternative to animal-derived feeder layers for growing hESCs.

Key words: Human embryonic stem cells (hESCs); Human feeder layer; Human menstrual blood-derived mesenchymal cells; Undifferentiated state

Received December 3, 2012; final acceptance January 31, 2014. Online prepub date: March 3, 2014.
Address correspondence to Regina Coeli dos Santos Goldenberg, Instituto de Biofisica Carlos Chagas FilhoUniversidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho 373 Bloco G - Sala G2-053, Rio de Janeiro, RJ 21941-902, Brazil. Tel: +55 21 2562 6559; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 7, pp. 37–47, 2014
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DOI: http://dx.doi.org/10.3727/215517914X680047
Copyright ©
2014 Cognizant Comm. Corp.
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Preculturing Islets With Adipose-Derived Mesenchymal Stromal Cells Is an Effective Strategy for Improving Transplantation Efficiency at the Clinically Preferred Intraportal Site

Chloe L. Rackham,1 Paramjeet K. Dhadda,1 Aurelie M. Le Lay, Aileen J. F. King, and Peter M. Jones

Diabetes Research Group, Division of Diabetes and Nutritional Sciences, Faculty of Life Sciences and Medicine, King’s College London, London, UK

We have recently shown that preculturing islets with kidney-derived mesenchymal stromal cells (MSCs) improves transplantation outcome in streptozotocin-diabetic mice implanted with a minimal mass of islets beneath the kidney capsule. In the present study, we have extended our previous observations to investigate whether preculturing islets with MSCs can also be used to enhance islet function at the clinically used intraportal site. We have used MSCs derived from adipose tissue, which are more readily accessible than alternative sources in human subjects and can be expanded to clinically efficacious numbers, to preculture islets throughout this study. The in vivo efficacy of grafts consisting of islets precultured alone or with MSCs was tested using asyngeneic streptozotocin-diabetic minimal islet mass model at the clinically relevant intraportal site. Blood glucose concentrations were monitored for 1 month. The vascularization of islets precultured alone or with MSCs was investigated both in vitro and in vivo, using immunohistochemistry. Islet insulin content was measured by radioimmunoassay. The effect of preculturing islets with MSCs on islet function in vitro was investigated using static incubation assays. There was no beneficial angiogenic influence of MSC preculture, as demonstrated by the comparable vascularization of islets precultured alone or with MSCs, both in vitro after 3 days and in vivo 1 month after islet transplantation. However, the in vitro insulin secretory capacity of MSC precultured islets was superior to that of islets precultured alone. In vivo, this was associated with improved glycemia at 7, 14, 21, and 28 days posttransplantation, in recipients of MSC precultured islets compared to islets precultured alone. The area of individual islets within the graft-bearing liver was significantly higher in recipients of MSC precultured islets compared to islets precultured alone. Our experimental studies suggest that preculturing islets with MSCs represents a favorable strategy for improving the efficiency of clinical islet transplantation.

Key words: Diabetes; Islet transplantation; Mesenchymal stromal cells (MSCs); Islet culture

Received November 01, 2013; final acceptance March 12, 2014. Online prepub date: March 24, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Chloe L. Rackham, Diabetes Research Group, Division of Diabetes and Nutritional Sciences, Faculty of Life Sciences and Medicine, King’s College London, London SE1 1UL, UK. Tel: +44 207 8486279; Fax: +44 207 8486280; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it