Peptides extracted from vero cell cultures overcome the blastocyst block of mouse embryos in a serum-free medium

Hsin Fu Chen, Hong Nerng Ho, Shee Uan Chen, Kuang Han Chao, Heng Ru Lin, Su Cheng Huang, Tzu Yao Lee, Yu Shih Yang

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Purpose: The aim of this work was to evaluate the effect of a Vero cell coculture system on the development of mouse embryos. Methods: Mouse embryos were randomly divided and cultured in human tubal fluid (HTF) medium with/without Vero cell monolayers, conditioned medium (CM) obtained from Vero cell cultures, and HTF medium supplemented with peptides extracted from CM. The concentrated CM was examined by SDS/PAGE. Results: The development of mouse embryos was blocked at the blastocyst stage in pure HTF medium (1.4% hatching at day 5). This "blastocyst block≓ was overcome by coculture with Vero cell monolayers (48.1% hatching at day 5; 1.4 vs 48.1%; P<0.001). CM and the addition of 5% fetal bovine serum (24.1 and 34.9% hatching, respectively, at day 5) were also able to enhance the process of hatching. In the other experiment, the addition of peptides extracted from Vero cell cultures also overcame the blastocyst block (12.5%) compared with pure HTF medium (2.1%) (P<0.05). Electrophoretic separation revealed several classes of polypeptides consistently secreted into CM obtained from Vero cell cultures. Most peptides occurred in the M r range between 6.5 kd and 35.9 kd. Conclusion: A developmental block (blastocyst block) of mouse embryos in a serum- and protein-free medium (HTF) was discovered in this study. This block was effectively overcome by HTF plus serum and coculture with Vero cell monolayers and also by the peptides extracted from Vero cell-conditioned medium. We speculate that certain factors secreted or converted by Vero cells may be critical in hatching of mouse embryos. Further study of these factors may be helpful in delineating its mechanism.

Original languageEnglish
Pages (from-to)165-171
Number of pages7
JournalJournal of Assisted Reproduction and Genetics
Volume11
Issue number3
DOIs
Publication statusPublished - Mar 1 1994
Externally publishedYes

Fingerprint

Cercopithecus aethiops
Inbred ICR Mouse
Vero Cells
Serum-Free Culture Media
Blastocyst
Coculture Techniques
Cell Division
Culture Media
Embryonic Structures
Cell Culture Techniques
Molecular Weight
Conditioned Culture Medium
Peptides
Embryonic Development
Serum
Blood Proteins
Polyacrylamide Gel Electrophoresis

Keywords

  • blastocyst block
  • coculture
  • mouse embryo
  • peptides
  • Vero cell

ASJC Scopus subject areas

  • Reproductive Medicine
  • Genetics
  • Obstetrics and Gynaecology
  • Developmental Biology
  • Genetics(clinical)

Cite this

Peptides extracted from vero cell cultures overcome the blastocyst block of mouse embryos in a serum-free medium. / Chen, Hsin Fu; Ho, Hong Nerng; Chen, Shee Uan; Chao, Kuang Han; Lin, Heng Ru; Huang, Su Cheng; Lee, Tzu Yao; Yang, Yu Shih.

In: Journal of Assisted Reproduction and Genetics, Vol. 11, No. 3, 01.03.1994, p. 165-171.

Research output: Contribution to journalArticle

Chen, Hsin Fu ; Ho, Hong Nerng ; Chen, Shee Uan ; Chao, Kuang Han ; Lin, Heng Ru ; Huang, Su Cheng ; Lee, Tzu Yao ; Yang, Yu Shih. / Peptides extracted from vero cell cultures overcome the blastocyst block of mouse embryos in a serum-free medium. In: Journal of Assisted Reproduction and Genetics. 1994 ; Vol. 11, No. 3. pp. 165-171.
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AU - Ho, Hong Nerng

AU - Chen, Shee Uan

AU - Chao, Kuang Han

AU - Lin, Heng Ru

AU - Huang, Su Cheng

AU - Lee, Tzu Yao

AU - Yang, Yu Shih

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AB - Purpose: The aim of this work was to evaluate the effect of a Vero cell coculture system on the development of mouse embryos. Methods: Mouse embryos were randomly divided and cultured in human tubal fluid (HTF) medium with/without Vero cell monolayers, conditioned medium (CM) obtained from Vero cell cultures, and HTF medium supplemented with peptides extracted from CM. The concentrated CM was examined by SDS/PAGE. Results: The development of mouse embryos was blocked at the blastocyst stage in pure HTF medium (1.4% hatching at day 5). This "blastocyst block≓ was overcome by coculture with Vero cell monolayers (48.1% hatching at day 5; 1.4 vs 48.1%; P<0.001). CM and the addition of 5% fetal bovine serum (24.1 and 34.9% hatching, respectively, at day 5) were also able to enhance the process of hatching. In the other experiment, the addition of peptides extracted from Vero cell cultures also overcame the blastocyst block (12.5%) compared with pure HTF medium (2.1%) (P<0.05). Electrophoretic separation revealed several classes of polypeptides consistently secreted into CM obtained from Vero cell cultures. Most peptides occurred in the M r range between 6.5 kd and 35.9 kd. Conclusion: A developmental block (blastocyst block) of mouse embryos in a serum- and protein-free medium (HTF) was discovered in this study. This block was effectively overcome by HTF plus serum and coculture with Vero cell monolayers and also by the peptides extracted from Vero cell-conditioned medium. We speculate that certain factors secreted or converted by Vero cells may be critical in hatching of mouse embryos. Further study of these factors may be helpful in delineating its mechanism.

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