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

被引：20

作者：

Dick, Emily ^{[1
]}

Matsa, Elena ^{[1
]}

Bispham, Jayson ^{[1
]}

Reza, Mojgan ^{[2
,3
]}

Guglieri, Michela ^{[2
,3
]}

Staniforth, Andrew ^{[4
]}

Watson, Sue ^{[5
]}

Kumari, Rajendra ^{[5
]}

Lochmueller, Hanns ^{[2
,3
]}

Young, Lorraine ^{[1
]}

Darling, David ^{[6
]}

Denning, Chris ^{[1
]}

机构：

[1] Univ Nottingham, Ctr Biomol Sci, Wolfson Ctr Stem Cells Tissue Engn & Modelling ST, Nottingham NG7 2RD, England

[2] Newcastle Univ, NE England Stem Cell Inst, Int Ctr Life, Newcastle Upon Tyne NE1 3BZ, Tyne & Wear, England

[3] Newcastle Univ, Inst Human Genet, Int Ctr Life, Newcastle Upon Tyne NE1 3BZ, Tyne & Wear, England

[4] Queens Med Ctr, Dept Cardiovasc Med, Nottingham NG7 2UH, England

[5] Univ Nottingham, Queens Med Ctr, Sch Clin Sci, Div Preclin Oncol, Nottingham NG7 2UH, England

[6] Kings Coll London, Guys Kings & St Thomas Sch Med, Rayne Inst, Dept Haematol Med, London SE5 9NU, England

来源：

STEM CELL RESEARCH | 2011年 / 6卷 / 02期

基金：

英国医学研究理事会; 英国生物技术与生命科学研究理事会;

关键词：

CARDIOMYOCYTE DIFFERENTIATION; HUMAN FIBROBLASTS; GENERATION; INDUCTION;

D O I：

10.1016/j.scr.2010.10.002

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：158 / 167

页数：10

共 38 条

[1] Variegation and silencing in a lentiviral-based murine transgenic model [J].