| Evaluation of Vero Cell Co-culture System for Mouse Embryo Development. |
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E K Lee, Y J Choi, Y B Kim |
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| 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. |
| Key Words:
Mouse embryos, Vero cell coculture system, In-vitro fertilization |
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