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Two new protocols to enhance the production and isolation of human induced pluripotent stem cell lines

Lookup NU author(s): Dr Mojgan Reza, Dr Michela GuglieriORCiD, Professor Hanns Lochmuller

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Abstract

There are two critical stages in the retroviral reprogramming of somatic cells to produce human induced pluripotent stem cell (hiPSC) lines. One is the production of high titer virus required to reprogram somatic cells; the other is identification of true hiPSC colonies from heterogeneous cell populations, and their isolation and expansion to generate a sustainable, pluripotent stem cell line. Here we describe simple, time-saving methods to address the current difficulties at these two critical junctures. First, we have developed a method to increase the number of infectious viral units 600-fold. Second, we have developed a TRA-1-81-based positive selection column method for isolating "true" hiPSCs from the heterogeneous cell populations, which overcomes the labor-intensive and highly subjective method of manual selection of hiPSC colonies. We have used these techniques to produce 8 hiPSC lines from human fibroblasts and we believe that they are of considerable utility to researchers in the hiPSC field. (c) 2010 Elsevier B.V. All rights reserved.


Publication metadata

Author(s): Dick E, Matsa E, Bispham J, Reza M, Guglieri M, Staniforth A, Watson S, Kumari R, Lochmuller H, Young L, Darling D, Denning C

Publication type: Article

Publication status: Published

Journal: Stem Cell Research

Year: 2011

Volume: 6

Issue: 2

Pages: 158-167

Print publication date: 01/03/2011

ISSN (print): 1873-5061

ISSN (electronic):

Publisher: Elsevier BV

URL: http://dx.doi.org/10.1016/j.scr.2010.10.002

DOI: 10.1016/j.scr.2010.10.002


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