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Korean Journal of Fertility and Sterility 2002;29(1):13-20.
Published online March 1, 2002.
The Study on Vitrification and Ultrarapid Thawing of Human Embryonic Stem Cells.
Shin Yong Moon, Yong Bin Park, Hee Sun Kim, Ki Chung Sung, Sun Kyung Oh, Dae Woo Chun, Chang Suk Suh, Young Min Choi, Jung Gu Kim, Jin Yong Lee, Seok Hyun Kim
1Department of Obstetrics and Gynecology, College of MedicineSeoul National University, Seoul, Korea.
2Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea.
Abstract
OBJECTIVE
This study was carried out to establish the effectiveness of the vitrification method and the optimal cryoprotectants in the cryopreservation of human embryonic stem cells (ESC). MATERIALS AND METHODS: Human ESC clumps established at Seoul National University Hospital (SNUhES 1) were cryopreserved with the vitrification method using the EM grid. EDS and EFS40 were used as vitrification solutions. RESULTS: Between the EDS and EFS40 groups, there was no significant difference in the recovery rate after cryopreservation of human ESC. The formation rates of ESC colonies in the vitrified groups were significantly lower than those in the control ESC group (p<0.05, p<0.05). In addition, the formation rate of ESC colonies in the EDS group was significantly higher than that in the EFS40 group (p<0.05). The ESC colonies in the vitrified groups were significantly smaller after culture duration of 2 and 4 days, respectively, compared with the control ESC group (p<0.01, p<0.05). However, these effects could be reduced to nonsignificant level by the additional culture of ESC colonies. The vitrified human ESC retained the properties of pluripotent cells, including the expression of cell surface markers for the undifferentiated cells such as alkaline phosphatase and SSEA-4 (stage-specific embryonic antigen-4), and the expression of transcription factor Oct-4 (octamer-binding transcription factor-4), and the normal karyotype. CONCLUSION: The vitrification method using the EM grid and EDS solution was confirmed to be very effective for the cryopreservation of human ESC.
Key Words: Human embryonic stem cells (ESC); Cryopreservation; Vitrification; EDS solution; EFS40 solution; EM grid


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