Evaluation of unrestricted somatic stem cells as a feeder layer to support undifferentiated embryonic stem cells |
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| Authors: | Saeed Heidari Keshel Masoud Soleimani Mostafa Rezaei Tavirani Maryam Ebrahimi Reza Raeisossadati Hemad Yasaei Danial Afsharzadeh Mahmoud Jabbarvand Behroz Amir Atashi Saeid Amanpour Ahad Khoshzaban Reza Roozafzoon Gholam Reza Behrouzi |
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| Institution: | 1. Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran;2. Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran;3. Tissue Engineering Department, School of Advanced Medical Technology, Tehran University of Medical Science, Tehran, Iran;4. Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran;5. Institute of Cancer Genetics and Pharmacogenomics, Division of Biosciences, School of Health Sciences and Social Care, Brunel University, Uxbridge, Middlesex, UK;6. Universitat Politecnica de Catalunua, Department of Agribusiness, Biotechnology, Barcelona, Spain;7. Faculty of Medicine, Cancer Institute, Tehran University of Medical Sciences, Imam Khomeini Hospital, Tehran, Iran |
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| Abstract: | The use of unrestricted somatic stem cells (USSCs) holds great promise for future clinical applications. Conventionally, mouse embryonic fibroblasts (MEFs) or other animal‐based feeder layers are used to support embryonic stem cell (ESC) growth; the use of such feeder cells increases the risk of retroviral and other pathogenic infection in clinical trials. Implementation of a human‐based feeder layer, such as hUSSCs that are isolated from human sources, lowers such risks. Isolated cord blood USSCs derived from various donors were used as a novel, supportive feeder layer for growth of C4mES cells (Royan C4 ESCs). Complete cellular characterization using immunocytochemical and flow cytometric methods were performed on murine ESCs (mESCs) and hUSSCs. mESCs cultured on hUSSCs showed similar cellular morphology and presented the same cell markers of undifferentiated mESC as would have been observed in mESCs grown on MEFs. Our data revealed these cells had negative expression of Stat3, Sox2, and Fgf4 genes while showing positive expression for Pou5f1, Nanog, Rex1, Brachyury, Lif, Lifr, Tert, B2m, and Bmp4 genes. Moreover, mESCs cultured on hUSSCs exhibited proven differentiation potential to germ cell layers showing normal karyotype. The major advantage of hUSSCs is their ability to be continuously cultured for at least 50 passages. We have also found that hUSSCs have the potential to provide ESC support from the early moments of isolation. Further study of hUSSC as a novel human feeder layer may lead to their incorporation into clinical methods, making them a vital part of the application of human ESCs in clinical cell therapy. Mol. Reprod. Dev. 79: 709–718, 2012. © 2012 Wiley Periodicals, Inc. |
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