Derivation of Clinical-Grade Induced Pluripotent Stem Cell Lines from Erythroid Progenitor Cells in Xenofree Conditions.

Autor:	Singh G; Centre for Stem Cell Research (A Unit of inStem, Bengaluru), Christian Medical College, Vellore, India., Manian KV; Centre for Stem Cell Research (A Unit of inStem, Bengaluru), Christian Medical College, Vellore, India., Premkumar C; Centre for Stem Cell Research (A Unit of inStem, Bengaluru), Christian Medical College, Vellore, India., Srivastava A; Centre for Stem Cell Research (A Unit of inStem, Bengaluru), Christian Medical College, Vellore, India.; Department of Haematology, Christian Medical College, Vellore, India., Daniel D; Department of Transfusion Medicine, Christian Medical College, Vellore, India., Velayudhan SR; Centre for Stem Cell Research (A Unit of inStem, Bengaluru), Christian Medical College, Vellore, India. rvshaji@cmcvellore.ac.in.; Department of Haematology, Christian Medical College, Vellore, India. rvshaji@cmcvellore.ac.in.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2454, pp. 775-789.
DOI:	10.1007/7651_2021_349
Abstrakt:	One of the major hurdles in realizing the therapeutic potential of human-induced pluripotent stem cells (iPSC) is the generation of clinical-grade iPSC lines and their differentiated progenies for preclinical and clinical applications. Therefore, there is a need to have standardized protocols for efficient generation of clinical-grade iPSC lines from easily accessible somatic cells in feeder-free, xenofree GMP grade culture conditions without genomic integration of the reprogramming factors. Here, we provide a detailed protocol for expansion of erythroid progenitor cells from peripheral blood mononuclear cells (PBMNC) and generation of iPSC lines in feeder-free and xenofree culture conditions from these cells by using GMP grade reagents. With this optimized protocol, clinical-grade iPSC lines can be derived from erythroid progenitor cells expanded from peripheral blood, which is easy-to-access, minimally invasive, and can be obtained from any donors. It will have implications in developing a large number of iPSC lines from individual healthy donors, diseased patients, or donors with homozygous human leukocyte antigen (HLA) for "haplobanking." (© 2021. Springer Science+Business Media, LLC.)
Databáze:	MEDLINE
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