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Differentiation of definitive endoderm from human induced pluripotent stem cells on hMSCs feeder in a defined medium

Jaafarpour, Z. and Soleimani, M. and Hosseinkhani, S. and Karimi, M.H. and Yaghmaei, P. and Mobarra, N. and Geramizadeh, B. (2016) Differentiation of definitive endoderm from human induced pluripotent stem cells on hMSCs feeder in a defined medium. Avicenna Journal of Medical Biotechnology, 8 (1). pp. 2-8.

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Abstract

Background: The Definitive Endoderm (DE) differentiation using the undefined media and non-human feeders can cause contaminations in the generated cells for therapeutic applications. Therefore, generating safer and more appropriate DE cells is needed. This study compared five different methods to establish an appropriate method for inducing an efficient DE differentiation from Human Induced Pluripotent Stem Cells (hiPSCs) on an appropriate feeder in a more defined medium. Methods: Human Induced Pluripotent Stem Cells (hiPSCs) were cultured on inactivated feeders. Passaged hiPSCs, without feeder, were incubated for three days with Activin-A and different endodermal differentiation media including 1-FBS, 2-B27, 3- ITS and albumin fraction-V, 4-B27 and ITS and 5-like the third medium. The feeder cells in the first four methods were Mouse Embryonic Fibroblasts (MEFs) and in the fifth method were human adult bone marrow Mesenchymal Stem Cells (hMSCs). DE markers FOXA2, SOX17 and CXCR4 and also pluripotency marker OCT4 were evaluated using qRT-PCR, as well as FOXA2 by the immunocytochemistry. Results: QRT-PCR analysis showed that after three days, the expression levels of DE and pluripotency markers in the differentiated hiPSCs among all five groups did not have any significant differences. Similarly, the immunocytochemistry analysis demonstrated that the differentiated hiPSCs expressed FOXA2, with no significant differences. Conclusion: Despite this similarity in the results, the third differentiation medium has more defined and cost effective components. Furthermore, hMSC, a human feeder, is safer than MEF. Therefore, the fifth method is preferable among other DE differentiation methods and can serve as a fundamental method helping the development of regenerative medicine. © 2016, Avicenna Journal of Medical Biotechnology. All rights reserved.

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: chemokine receptor CXCR4; hepatocyte nuclear factor 3beta; octamer transcription factor 4; transcription factor Sox17, animal cell; Article; bone marrow cell; cell differentiation; controlled study; culture medium; definitive Endoderm; endoderm; feeder cell; female; human; human cell; immunocytochemistry; mesenchymal stem cell; mouse; nonhuman; pluripotent stem cell; protein expression; real time polymerase chain reaction
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 07 Sep 2016 08:56
Last Modified: 19 Sep 2016 10:35
URI: http://eprints.goums.ac.ir/id/eprint/4635

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