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Single cell transcriptional profiling reveals heterogeneity of human induced pluripotent stem cells
Kazim H. Narsinh, … , Robert C. Robbins, Joseph C. Wu
Kazim H. Narsinh, … , Robert C. Robbins, Joseph C. Wu
Published February 7, 2011
Citation Information: J Clin Invest. 2011;121(3):1217-1221. https://doi.org/10.1172/JCI44635.
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Brief Report Stem cells

Single cell transcriptional profiling reveals heterogeneity of human induced pluripotent stem cells

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Abstract

Human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) are promising candidate cell sources for regenerative medicine. However, despite the common ability of hiPSCs and hESCs to differentiate into all 3 germ layers, their functional equivalence at the single cell level remains to be demonstrated. Moreover, single cell heterogeneity amongst stem cell populations may underlie important cell fate decisions. Here, we used single cell analysis to resolve the gene expression profiles of 362 hiPSCs and hESCs for an array of 42 genes that characterize the pluripotent and differentiated states. Comparison between single hESCs and single hiPSCs revealed markedly more heterogeneity in gene expression levels in the hiPSCs, suggesting that hiPSCs occupy an alternate, less stable pluripotent state. hiPSCs also displayed slower growth kinetics and impaired directed differentiation as compared with hESCs. Our results suggest that caution should be exercised before assuming that hiPSCs occupy a pluripotent state equivalent to that of hESCs, particularly when producing differentiated cells for regenerative medicine aims.

Authors

Kazim H. Narsinh, Ning Sun, Veronica Sanchez-Freire, Andrew S. Lee, Patricia Almeida, Shijun Hu, Taha Jan, Kitchener D. Wilson, Denise Leong, Jarrett Rosenberg, Mylene Yao, Robert C. Robbins, Joseph C. Wu

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Figure 1

Gene expression profiling of 282 single hESCs and hiPSCs.

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Gene expression profiling of 282 single hESCs and hiPSCs.
(A) An array s...
(A) An array showing that pluripotency (top) and lineage (bottom) marker transcripts are differentially expressed in pluripotent stem cell types (H7, H9, HES2, hiPSC1–4) versus somatic cell types (IMR90 and hASCs). (B) Comparison of the Ct values obtained by single cell qRT-PCR shows that pluripotency transcript levels are equivalent when considering the cell populations as a whole (geometric mean ± SD). (C) Population distribution plots (horizontal axis represents Ct value; vertical axis represents percentage of total cell population) reveal that single hiPSCs (white bars) display considerable heterogeneity in transcript expression levels in comparison with that of single hESCs (black bars).
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