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Human Embryonic Stem Cells (hESCs) Cultured Under Distinctive Feeder-Free Culture Conditions Display Global Gene Expression Patterns Similar to hESCs from Feeder-Dependent Culture Conditions

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Abstract

Human embryonic stem cell (hESC)–based assay systems and genetically modified hESCs are very useful tools for screening drugs that regulate stemness and differentiation and for studying the molecular mechanisms involved in hESC fate determination. For these types of studies, feeder cell–dependent cultures of hESCs are often problematic because the physiology of the feeder cells is perturbed by the drug treatments or genetic modifications, which potentially obscures research outcomes. In this study, we evaluated three commonly used feeder-free culture conditions to determine whether they supported the undifferentiated growth of hESCs and to determine whether the hESCs grown in these conditions displayed gene expression patterns that were similar to the expression patterns of feeder cell-dependent hESCs. Our results demonstrate that hESCs grown in the three feeder-free conditions expressed undifferentiation marker genes as strongly as hESCs that were grown in the feeder-dependent cultures. Furthermore, genome-wide gene expression profiles indicated that the gene expression patterns of hESCs that were grown under feeder-free or feeder-dependent culture conditions were highly similar. These results indicate that the feeder-free culture conditions support the undifferentiated growth of hESCs as effectively as the feeder-dependent culture conditions. Therefore, feeder-free culture conditions are potentially suitable for drug screening and for the genetic manipulation of hESCs in basic research.

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Acknowledgments

This research was supported by grants (SC5170) from the Stem Cell Research Center of the 21st Century Frontier Research Program, Bio R&D Program, and the World Class University program through the National Research Foundation of Korea (R31-2008-000-10086-0), all funded by the Ministry of Education, Science and Technology, Republic of Korea.

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Authors and Affiliations

  1. CHA Stem Cell Institute, Department of Biomedical Science, CHA University College of Medicine, Seoul, 135-081, South Korea

    Tae-Min Yoon, Bomi Chang, Hyeung-Taek Kim, Joo-Hyun Jee & Dong-Youn Hwang

  2. Department of Physiology, Yonsei University College of Medicine, Seoul, 120-752, South Korea

    Dong-Wook Kim

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  1. Tae-Min Yoon
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  2. Bomi Chang
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Corresponding author

Correspondence to Dong-Youn Hwang.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary Figure 1

Images of higher magnifications (×200) are presented for clear visualization of the individual cell morphologies of the CHA6-hESCs and H9-hESCs that have been grown in different feeder-free conditions. Both cell lines were immunostained with antibody against Oct4. Scale bars: 100 μm. (GIF 382 kb)

High Resolution Image (TIFF 1.71 mb)

Supplementary Table 1

Forty-six of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in MEF feeder-dependent cultures were compared to the gene levels in the hESCs grown in the Matrigel/mTeSR-1 culture condition. Seventeen specific gene-level probes that are also listed in Supplementary Tables 2 and 3 are marked in yellow. (GIF 74 kb)

High Resolution Image (TIFF 620 kb)

Supplementary Table 2

Forty-seven of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in MEF feeder-dependent cultures were compared to the gene levels in the hESCs grown in the vitronectin/mTeSR-1 culture condition. Seventeen specific gene-level probes that are also listed in Supplementary Tables 1 and 3 are marked in yellow. (GIF 251 kb)

High Resolution Image (TIFF 147 kb)

Supplementary Table 3

Forty-six of the most significantly upregulated gene-level probes in the CHA6-hESCs that were grown in MEF feeder-dependent cultures were compared to the gene levels in the hESCs that were grown in the CELLstart/STEMPRO culture condition. Seventeen specific gene-level probes that are also listed in Supplementary Tables 1 and 2 are marked in yellow. (GIF 244 kb)

High Resolution Image (TIFF 146 kb)

Supplementary Table 4

A total of 12 gene-level probes that are expressed at higher levels in the CHA6-hESCs grown in the Matrigel/mTeSR-1 condition than in the hESCs grown in the vitronectin/mTeSR-1 culture condition. The identities and names of all of the gene-level probes are not yet assigned. (GIF 119 kb)

High Resolution Image (TIFF 82 kb)

Supplementary Table 5

Forty-three of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the Matrigel/mTeSR-1 condition were compared to the gene levels in the hESCs grown in the CELLstart/STEMPRO culture condition. Eight of the specific gene-level probes that are also listed in Supplementary Table 6 are marked in orange. (GIF 243 kb)

High Resolution Image (TIFF 137 kb)

Supplementary Table 6

Forty-two of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the Matrigel/mTeSR-1 condition were compared to the gene levels in the hESCs grown in the MEF feeder-dependent culture condition. Eight of the specific gene-level probes that are also listed in Supplementary Table 5 are marked in orange. (GIF 225 kb)

High Resolution Image (TIFF 138 kb)

Supplementary Table 7

Twenty-two gene-level probes that are expressed at higher levels in the CHA6-hESCs grown in the vitronectin/mTeSR-1 condition than in the hESCs grown in the Matrigel/mTeSR-1 culture condition. The identities and names of all but three of the gene-level probes are not assigned. (GIF 159 kb)

High Resolution Image (TIFF 110 kb)

Supplementary Table 8

Forty-four of the most differentially expressed gene-level probes between the CHA6-hESCs grown in the vitronectin/mTeSR-1 conditions and the hESCs grown in the MEF feeder-dependent culture condition. Five specific gene-level probes that are also listed in Supplementary Table 9 are marked in green. (GIF 221 kb)

High Resolution Image (TIFF 139 kb)

Supplementary Table 9

Thirty-two of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the vitronectin/mTeSR-1 condition were compared to the gene levels in the hESCs grown in the CELLstart/STEMPRO culture condition. Five specific gene-level probes that are also listed in Supplementary Table 8 are marked in green. (GIF 190 kb)

High Resolution Image (TIFF 120 kb)

Supplementary Table 10

Forty-five of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the CELLstart/STEMPRO condition were compared to the gene levels in the hESCs grown in the MEF feeder-dependent culture condition. Five specific gene-level probes that are also listed in Supplementary Tables 11 and 12 are marked in gray. (GIF 222 kb)

High Resolution Image (TIFF 155 kb)

Supplementary Table 11

Twenty-three of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the CELLstart/STEMPRO condition were compared to the gene levels in the hESCs grown in the Matrigel/mTeSR-1 culture condition. Five specific gene-level probes that are also listed in Supplementary Tables 10 and 12 are marked in gray. (GIF 158 kb)

High Resolution Image (TIFF 107 kb)

Supplementary Table 12

Twenty-eight of the most significantly upregulated gene-level probes in the CHA6-hESCs grown in the CELLstart/STEMPRO condition were compared to the gene levels in the hESCs grown in the vitronectin/mTeSR-1 culture condition. Five specific gene-level probes that are also listed in Supplementary Tables 11 and 12 are marked in gray. (GIF 159 kb)

High Resolution Image (TIFF 110 kb)

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Yoon, TM., Chang, B., Kim, HT. et al. Human Embryonic Stem Cells (hESCs) Cultured Under Distinctive Feeder-Free Culture Conditions Display Global Gene Expression Patterns Similar to hESCs from Feeder-Dependent Culture Conditions. Stem Cell Rev and Rep 6, 425–437 (2010). https://doi.org/10.1007/s12015-010-9158-x

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