Derivation, characterization and retinal differentiation of induced pluripotent stem cells.

dc.contributor.authorMekala, Subba Rao
dc.contributor.authorVauhini, Vasundhara
dc.contributor.authorNagarajan, Usha
dc.contributor.authorMaddileti, Savitri
dc.contributor.authorGaddipati, Subhash
dc.contributor.authorMariappan, Indumathi
dc.date.accessioned2015-08-12T07:13:06Z
dc.date.available2015-08-12T07:13:06Z
dc.date.issued2013-03
dc.description.abstractMillions of people world over suffer visual disability due to retinal dystrophies which can be age-related or a genetic disorder resulting in gradual degeneration of the retinal pigmented epithelial (RPE) cells and photoreceptors. Therefore, cell replacement therapy offers a great promise in treating such diseases. Since the adult retina does not harbour any stem cells, alternative stem cell sources like the embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer a great promise for generating different cell types of the retina. Here, we report the derivation of four iPSC lines from mouse embryonic fibroblasts (MEFs) using a cocktail of recombinant retroviruses carrying the genes for Oct4, Sox2, Klf4 and cMyc. The iPS clone MEF-4F3 was further characterized for stemness marker expression and stable reprogramming by immunocytochemistry, FACS and RT-PCR analysis. Methylation analysis of the nanog promoter confirmed the reprogrammed epigenetic state. Pluripotency was confirmed by embryoid body (EB) formation and lineage-specific marker expression. Also, upon retinal differentiation, patches of pigmented cells with typical cobble-stone phenotype similar to RPE cells are generated within 6 weeks and they expressed ZO-1 (tight junction protein), RPE65 and bestrophin (mature RPE markers) and showed phagocytic activity by the uptake of fluorescent latex beads.en_US
dc.identifier.citationMekala Subba Rao, Vauhini Vasundhara, Nagarajan Usha, Maddileti Savitri, Gaddipati Subhash, Mariappan Indumathi. Derivation, characterization and retinal differentiation of induced pluripotent stem cells. Journal of Biosciences. 2013 Mar; 38(1): 123-134.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/161799
dc.language.isoenen_US
dc.source.urihttps://www.ias.ac.in/jbiosci/mar2013/123.pdfen_US
dc.subjectInduced pluripotent stem cells (iPSCs)en_US
dc.subjectmouse embryonic fibroblasts (MEFs)en_US
dc.subjectretinal pigmented epithelium (RPE)en_US
dc.titleDerivation, characterization and retinal differentiation of induced pluripotent stem cells.en_US
dc.typeArticleen_US
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