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Derivation and long-term culture of human parthenogenetic embryonic stem cells using human foreskin feeders

  • Stem cell biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Feeder cells from animals raise considerable concern for contamination because they are directly in contact with embryonic stem cells.

Methods

To address this issue we collected discarded foreskin tissue and prepared a fibroblast cell line. We transferred one parthenogenetic blastocyst on to these feeder cells, and later observed outgrowth. By this approach, we were able to derive a human parthenogenetic embryonic stem cell line successfully.

Results

The embryonic stem cells had normal morphology, expressed all expected cell surface markers, could differentiate to embryonic bodies upon culture in vitro, and differentiated further to derivatives of all three germ layers.

Conclusion

This study indicates that homologous human fibroblasts can be used as feeder cells to support not only the propagation, but also the derivation of ES cells, and this should facilitate studies of therapeutic cloning for research and clinical applications.

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Acknowledgements

We are grateful to Michelle Yu for the invaluable collaboration in the writing and revision of the manuscript. This work was supported by grants from the Major State Basic Research Development Program of China (973 Program) (No. 2007CB948001) and the National High Technology Research and Development Program of China (863 Program) (No. 2006AA02A101).

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Authors and Affiliations

  1. Center for Reproductive Medicine, Tianjin Central Hospital for Obstetrics and Gynecology, Tianjin, 300052, People’s Republic of China

    Zhenyu Lu, Ruqiang Yao & Yunshan Zhang

  2. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 1st Beichen West Road, Chaoyang District, Beijing, 100101, People’s Republic of China

    Wanwan Zhu, Yang Yu & Qi Zhou

  3. Cell Therapy Center, Xuanwu Hospital, Capital Medical University, Beijing, 100053, People’s Republic of China

    Wanwan Zhu, Yunqian Guan & Yu Alex Zhang

  4. Tianjin Medical University, Tianjin, 300070, People’s Republic of China

    Zhenyu Lu & Dan Jin

Authors
  1. Zhenyu Lu
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  2. Wanwan Zhu
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  3. Yang Yu
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  9. Qi Zhou
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Corresponding author

Correspondence to Qi Zhou.

Additional information

Zhenyu Lu, Wanwan Zhu and Yang Yu contributed equally to this work.

Capsule One human embryonic stem cell line from parthenogenetic embryos was derived using human foreskin feeder cells and maintained normal characteristics during long term propagation.

Electronic Supplementary Material

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Supplement Table 1

PCR primers used to detect gene expression in human ES and EB cells (DOC 34 kb)

Supplement Fig. 1

Colony of line P-TJ (passage level 15) growing on the hFF feeder layer. Bar: 100 μm (GIF 218 kb)

High resolution image file (TIFF 3614 kb)

Supplement Fig. 2

RT-PCR analysis of the expression of pluripotent genes in line P-TJ (lanes 1–4). The hES cell line H9 was analyzed at the same time as a control (lanes 5–8). Lanes 1, 5: β-actin; lanes 2, 6: OCT4; lanes 3, 7: NANOG; lanes 4, 8: REX1 (GIF 42 kb)

High resolution image file (TIFF 57 kb)

Supplement Fig. 3

Embryoid bodies at day 4 in suspension culture conditions. Bar: 100 μm (GIF 128 kb)

High resolution image file (TIFF 219 kb)

Supplement Fig. 4

T-PCR analysis of the expression of selected imprinted genes. The human parthenogenetic embryonic stem cells expressed only maternal genes (lanes 1, 2), and not paternal genes (lanes 3, 4). The control hES cell line H9 expressed both maternal and paternal genes (lanes 5–8). Lanes 1, 5: H19; lanes 2, 6: UBE3A; lanes 3, 7: hRPNR; lanes 4, 8: IGF2 (GIF 27 kb)

High resolution image file (TIFF 195 kb)

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Lu, Z., Zhu, W., Yu, Y. et al. Derivation and long-term culture of human parthenogenetic embryonic stem cells using human foreskin feeders. J Assist Reprod Genet 27, 285–291 (2010). https://doi.org/10.1007/s10815-010-9408-5

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  • DOI: https://doi.org/10.1007/s10815-010-9408-5

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