Korean J Obstet Gynecol.  1999 May;42(5):1069-1073.

Evaluation of Vero Cell Co-culture System for Mouse Embryo Development

• Lee EK
• Choi YJ
• Kim YB

Abstract

OBJECTIVE
To demonstrate whether Vero cells and their conditioned media enhance mouse embryo development and to determine the mechanism of action by which these cells function.
MATERIALS AND METHODS
Vero cells can be commercially obtained in a frozen state. From the frozen cells, flasks were seeded at 2 to 3x106 cells, reaching confluent monolayer within 3days. Conditioned media were prepared from Vero cell monolayer following a 48 hours conditioning period in 1,000 microl of Ham`s F10 media in four well plate at 37degreeC and 5% CO2 in humidified air. 1-cell mouse embryos [Crl:CD:1[ICR]BR] were incubated on Vero cell monolayer with Ham`s F10 supplemented with 10%FBS, as well as in conditioned media at 37degreeC in an atmosphere of 5% CO2. The rate of development to the blastocyst stage from the 1-cell embryo was examined in all sets of experiments and compared with those cultured in control medium alone.
RESULTS
In early embryo development, significantly more 3-to 8-cell embryos developed in CM group than control group [36.6 versus 12.3% ; p=0.009], but there was no significant difference between Vero cell and CM group [46.3 versus 36.6% ; p=0.354]. In late embryo development, completion of hatching was higher in Vero cell group than in CM group [12.5 versus 8.1% ; p=0.020]. However, no difference was detected in blastocyst development between Vero cell and CM group[39.9 versus 40.5%; p=0.077].
CONCLUSION
This study suggested that [1] both Vero cell co-culture and conditioned medium supported development to blastocyst stage at a higher efficiency than control medium alone. [2] This is likely to be due to the secretion of embryotrophic factors rather than removal of substances inhibitory to development. [3] But the production of various growth factors by Vero cells may have been reduced during the brief medium-conditioning period[48 hours], which could account for the lowered hatching blastocyst formation in the conditioned medium.

Keyword

Mouse embryos; Vero cell coculture system; In-vitro fertilization

MeSH Terms

Animals
Atmosphere
Blastocyst
Coculture Techniques*
Culture Media, Conditioned
Embryonic Development*
Embryonic Structures*
Female
Intercellular Signaling Peptides and Proteins
Mice*
Pregnancy
Vero Cells*
Culture Media, Conditioned
Intercellular Signaling Peptides and Proteins