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Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

ABSTRACT: Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue-culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with Inter-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IαI-supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications.

ORGANISM(S): Homo sapiens

PROVIDER: GSE82103 | GEO | 2016/06/02

SECONDARY ACCESSION(S): PRJNA324010

REPOSITORIES: GEO

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Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

Project description:Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue-culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with Inter-Î±-inhibitor (IÎ±I), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IÎ±I efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IÎ±I-supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications. SNP genotyping with Illumina HumanOmniExpressExome-array version 8v1-2_A, encompassing >964.000 single nucleotide polymorphism (SNP)-markers, were performed by the SNP&SEQ technology Platform in Uppsala (www.genotyping.se), according to the manufacturerâ??s instructions. Each of the four hPS cell lines (i.e. H181, H207, HUES1 and K2C) were analyzed in three separate SNP-array experiments for the 12 samples described here. A first analysis was performed after 2-5 passages of growth for initial reference (4 samples), and subsequently, two analyses were performed per cell line after 16-21 passages in E8:VN or E8:IÎ±I culture (8 samples). The CN-calls of the autosomal chromosomes were illustrated using the Nexus-software.

2016-06-02 | E-GEOD-82103 | biostudies-arrayexpress

Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture.

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A stress-reduced passaging technique improves the viability of human pluripotent cells

Project description:Xeno-free culture systems have expanded the application of human pluripotent stem cells (PSCs). Here we describe an improved method for the subculture of human PSCs. The new method significantly facilitated the viability of human PSCs by lowering DNA damage and apoptosis, resulting in more efficient and reproducible downstream applications. Furthermore, the method did not alter the PSC characteristics after long-term culture and attenuated the growth advantage of abnormal subpopulations. This robust passaging method minimizes experimental error and improves the quality control of human PSC research and applications.

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Global transcriptional analysis of xeno-free and feeder-cultured human extended pluripotent stem cells by RNA-seq

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Chromatin accessibility of xeno-free and feeder-cultured human extended pluripotent stem cells by ATAC-seq

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Extracellular matrix-dependent generation of integration- and xeno-free iPS cells using a modified mRNA transfection method

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Analysis of Mast Cells in Hermansky-Pudlak Syndrome Type-1

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2016-07-01 | E-GEOD-83920 | biostudies-arrayexpress

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