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Surface Marker Epithelial Cell Adhesion Molecule and E-cadherin Facilitate the Identification and Selection of Induced Pluripotent Stem Cells

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Abstract

The derivation of induced pluripotent stem cells (iPSCs) requires not only efficient reprogramming methods, but also reliable markers for identification and purification of iPSCs. Here, we demonstrate that surface markers, epithelial cells adhesion molecule (EpCAM) and epithelial cadherin (E-cadherin) can be used for efficient identification and/or isolation of reprogrammed mouse iPSCs. By viral transduction of Oct4, Sox2, Klf4 and n- or c-Myc into mouse embryonic fibroblasts, we observed that the conventional mouse embryonic stem cell (mESC) markers, alkaline phosphatase (AP) and stage-specific embryonic antigen 1 (SSEA1), were expressed in incompletely reprogrammed cells that did not express all the exogenous reprogramming factors or failed to acquire pluripotent status even though exogenous reprogramming factors were expressed. EpCAM and E-cadherin, however, remained inactivated in these cells. Expression of EpCAM and E-cadherin correlated with the activation of Nanog and endogenous Oct4, and was only seen in the successfully reprogrammed iPSCs. Furthermore, purification of EpCAM-expressing cells at late reprogramming stage by FACS enriched the Nanog-expressing cell population suggesting the feasibility of selecting successful reprogrammed mouse iPSCs by EpCAM expression. We have thus identified new surface markers that can efficiently identify successfully reprogrammed iPSCs and provide an effective means for iPSC isolation.

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Acknowledgements

We are grateful to Shih-Chih Tsai for assistance in preparation of manuscript. This work was supported by an intramural grant from Academia Sinica, and Stem Cell Priority Grants (#97-3111-B-001-009 and 98-2811-B-001-022) from the National Science Council, Taiwan.

Disclosures

The authors indicate no potential conflicts of interest.

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Authors and Affiliations

  1. Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, College of Medicine and the Hospital, National Taiwan University, Taipei, Taiwan

    Hsin-Fu Chen & Hong-Nerng Ho

  2. Graduate Institute of Clinical Genomics, College of Medicine, National Taiwan University, Taipei, Taiwan

    Hsin-Fu Chen & Hong-Nerng Ho

  3. Institute of Cellular and Organismic Biology, Academia Sinica, No. 128, Sec. 2, Academia Road, Nankang, Taipei, Taiwan

    Wen-Chih Lee, Han-Chung Wu & Hung-Chih Kuo

  4. Genomics Research Center, Academia Sinica, Taipei, Taiwan

    Ching-Yu Chuang

  5. Division of Medical Research, National Taiwan University Hospital, Taipei, Taiwan

    Hsiang-Po Huang

  6. Institute of Chemistry, Academia Sinica, Taipei, Taiwan

    Yu-Ju Chen

  7. Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan

    Hung-Chih Kuo

  8. Institute of Biotechnology, National Taiwan Ocean University, Keelung, Taiwan

    Wen-Chih Lee & Hung-Chih Kuo

  9. Stem Cell Program, Genomics Research Center, Academia Sinica, No. 128, Sec. 2, Academia Road, Nankang, Taipei, Taiwan

    Hung-Chih Kuo

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  1. Hsin-Fu Chen
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Correspondence to Hung-Chih Kuo.

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Supplementary Table 1

(GIF 158 kb)

High resolution image file (TIFF 21897 kb)

Supplementary Fig. 1

Flow cytometry histogram of percentages of cells expressing SSEA1, EpCAM and E-Cadherin in MEFs, mESCs and mESC-like cells by lentiviral transduction. (GIF 188 kb)

High resolution image file (TIFF 17863 kb)

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Chen, HF., Chuang, CY., Lee, WC. et al. Surface Marker Epithelial Cell Adhesion Molecule and E-cadherin Facilitate the Identification and Selection of Induced Pluripotent Stem Cells. Stem Cell Rev and Rep 7, 722–735 (2011). https://doi.org/10.1007/s12015-011-9233-y

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