Obstet Gynecol Sci.  2013 Sep;56(5):320-329.

The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture

Affiliations
  • 1Department of Obstetrics and Gynecology, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon, Korea. oldfox@cnuh.co.kr
  • 2Fertility Center, Miz Woman's Hospital, Daejeon, Korea.
  • 3Infertility Clinic, Department of Obstetrics and Gynecology, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon, Korea.

Abstract


OBJECTIVE
This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos.
METHODS
A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05.
RESULTS
The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P < 0.05). The rates of embryonic development after 48 hours (morula< or =) and 96 hours (blastocyst< or =) were significantly higher in 20% SSS and 10% SPS than in 20% hFF supplementation (P < 0.05). And the rates of embryonic development after 96 hours (hatching blastocyst< or =) were significantly higher in 10% SPS (94.5%) than in 20% SSS (82.6%) and 20% hFF supplementation (68.5%). The rates of embryonic development according to storage period of the SF-VCM supplemented with 10% SPS showed no significant difference between control, 2 weeks and 4 weeks group. However developmental rate in 6 weeks storage group was significantly lower than other groups.
CONCLUSION
The rate of embryonic development after 96 hours (hatching blastocyst< or =) was significantly higher in SF-VCM supplemented with 10% SPS. And storage period of media up to 4 weeks did not affect on embryonic development.

Keyword

Coculture; Mouse embryo; Serum free culture media, Vero cells

MeSH Terms

Animals
Coculture Techniques
Culture Media, Conditioned
Eagles
Embryonic Development
Embryonic Structures
Female
Follicular Fluid
Humans
Mice
Pregnancy
Vero Cells
Culture Media, Conditioned

Figure

  • Fig. 1 Comparison between the effects of different supplementations on in vitro development of mouse embryos cultured in SF-VCM. SF-VCM, serum free Vero cell conditioned media; hFF, human follicular fluid; SSS, serum substitute supplement; SPS, serum protein substitute; BL, blastocyst. a)P<0.05 vs. 20% hFF; b)P<0.05 v.s 20% SSS.


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