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Stem cell factor/c-Kit signaling in in vitro cultures supports early mouse embryonic development by accelerating proliferation via a mechanism involving Akt-downstream genes

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

Abstract

Purpose

Stem cell factor (SCF)/c-Kit regulates the proliferation and survival of germ cells or stem cells; however, little is known about the role of SCF/c-Kit in pre-implantation embryo development.

Methods

Using exogenous SCF supplementation and c-Kit siRNA injection, we investigated the role and mechanism of SCF/c-Kit in pre-implantation mouse embryos.

Results

Addition of soluble SCF to the culture medium improved blastocyst formation. c-Kit gene silencing reduced the rate of blastocyst formation and delayed embryonic development. The number of proliferating cells in c-Kit gene-silenced blastocysts decreased, whereas the number of apoptotic cells in blastocysts obtained from both experimental and the control groups was not affected. RT-PCR, immunostaining and western blotting revealed that proliferation-related Akt downstream targets were substantially affected by c-Kit gene silencing.

Conclusion

SCF/c-Kit signaling through Akt downstream targets is likely involved in mediating the cleavage and proliferation of blastomeres during mouse pre-implantation embryogenesis.

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Acknowledgements

This research was supported by a grant (2009-0093821) from Priority Research Centers Program funded by the Ministry of Education, Science and Technology, Republic of Korea.

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Authors and Affiliations

  1. Fertility Center, CHA Gangnam Medical Center, CHA University, 606-5 Yeoksam-dong, Gangnam-gu, Seoul, 135-081, South Korea

    Jung Jin Lim, Jin Hee Eum, Jeoung Eun Lee, Eun Sun Kim, Hyung Min Chung, Tae Ki Yoon & Dong Ryul Lee

  2. Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 133–791, South Korea

    Jung Jin Lim & Kye-Seong Kim

  3. Department of Biomedical Science, College of Life Science, CHA University, Seoul, 135-081, South Korea

    Dong Ryul Lee

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  1. Jung Jin Lim
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Corresponding authors

Correspondence to Kye-Seong Kim or Dong Ryul Lee.

Additional information

Capsule

c-Kit siRNA inhibited development of preimplantation-stage mouse embryo and changed the mRNA expression of Akt target gene (upregulation of Tuberin and downregulation of mTOR). Tuberin normally inhibited mTOR which modulates cell proliferlation.

Jung Jin Lim and Jin Hee Eum contributed equally to this study.

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Supplemental Figure 1

Effect of SCF supplementation on the development of mouse one-cell embryos at 96 h post-hCG. The data are expressed as the mean±SEM. a,b Values within the same column with different superscripts are significantly different (P < 0.05). Note: KSOM (only): 139 zygotes, serum-free KSOM media, KSOM (100 ng/ml SCF): 138 zygotes, serum-free KSOM media with 100 ng/ml soluble SCF, KSOM (3 mg/ml BSA): 144 zygotes, KSOM media with 3 mg/ml BSA. Early Bla: early blastocyst (less than 50% cavity), Bla: blastocyst (greater than 50% cavity), Expanded Bla: expanded blastocyst, Hatching Bla: hatching blastocyst. (GIF 170 kb)

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Lim, J.J., Eum, J.H., Lee, J.E. et al. Stem cell factor/c-Kit signaling in in vitro cultures supports early mouse embryonic development by accelerating proliferation via a mechanism involving Akt-downstream genes. J Assist Reprod Genet 27, 619–627 (2010). https://doi.org/10.1007/s10815-010-9449-9

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  • DOI: https://doi.org/10.1007/s10815-010-9449-9

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