Early selection of human fibroblast-derived induced pluripotent stem cells

Abstract

Induced pluripotent stem cells (iPSC) are ‘artificial’ stem cells that can be obtained by reprogramming of somatic cells. iPSC display properties, typical for embryonic stem cells (ESC) in terms of morphology, proliferation, differentiation capacity, genomic and epigenomic states. The generation of iPSC from somatic cells not only reduces some of the problems, connected with ESC, but also provides a potential human in vitro model for disease modelling and drug development. The current techniques for the production of iPSC are ineffective and often incomplete. Оbtaining of extremely low amount of fully reprogrammed cells or iPSC is one of the current problems. The low efficiency of iPSC generation is a significant barrier for fast and accurate screening and makes the colony isolation time consuming and costly. In order to develop the full potential of iPSC technology, scientists are searching for new and more effective methods for generation and isolation of iPSC. Here, we report a method that could help for the isolation of iPSC cells in a short period after reprogramming. The method involves a selection of cells, based on the expression of OCT4 and a surface pluripotent marker TRA-1-60. Reprogrammed cells, isolated by this method, expressed pluripotent markers OCT4, SOX2, NANOG, KLF4 and TRA-1-60 and were able to differentiate into the three germ layers in vitro.

Keywords:

• cell reprogramming
• human-induced pluripotent stem cells
• human fibroblasts
• selection of iPSC

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by Bulgarian Ministry of Education and Science [grant number D01-4787/ReProForce FP-7- REGPOT-2009-1]; the European Social Fund and Operational Programme Human Resources Development (2007–2013) with co-funding from the Bulgarian Ministry of Education and Science [grant number BG051PO001-3.3.06-0059].