KMID : 0358220060330040265
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Korean Journal of Fertility and Sterility 2006 Volume.33 No. 4 p.265 ~ p.272
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Effect of the Isolation Method of Mouse Inner Cell Mass, Types of Feeder Cells and Treatment Time of Mitomycin C on the Formation Rate of ICM Colony
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Jang Ho-Jin
Ko Gyoung-Rae Kim Mi-Kyung Na Yong-Jin Lee Kyu-Sup
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Abstract
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Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony.
Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days.
Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions.
Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.
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KEYWORD
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ICM colony, Feeder layer, Isolation method, Mitomycin C
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