»ýÁã ³»¼¼Æ÷±«ÀÇ ºÐ¸®¹æ¹ý°ú ÁöÁö¼¼Æ÷ÀÇ Á¾·ù¿Í Mitomycin C ó¸® ½Ã°£ÀÌ ³»¼¼Æ÷±« Colony Çü¼º·ü¿¡ ¹ÌÄ¡´Â ¿µÇâ
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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ÀåÈ£Áø ( Jang Ho-Jin )
ºÎ»ê´ëÇб³ ÀÇ°ú´ëÇÐ »êºÎÀΰúÇб³½Ç
°í°æ·¡ ( Ko Gyoung-Rae )
ºÎ»ê´ëÇб³º´¿ø ºÒÀÓŬ¸®´Ð
±è¹Ì°æ ( Kim Mi-Kyung )
ºÎ»ê´ëÇб³º´¿ø ºÒÀÓŬ¸®´Ð
³ª¿ëÁø ( Na Yong-Jin )
ºÎ»ê´ëÇб³ ÀÇ°ú´ëÇÐ »êºÎÀΰúÇб³½Ç
À̱Լ· ( Lee Kyu-Sup )
ºÎ»ê´ëÇб³ ÀÇ°ú´ëÇÐ »êºÎÀΰúÇб³½Ç
KMID : 0358220060330040265
Abstract
¸ñ Àû: º» ¿¬±¸´Â »ýÁã Æ÷¹è±â ¹è¾Æ·ÎºÎÅÍ ³»¼¼Æ÷±«¸¦ ºÐ¸®ÇÏ´Â ¹æ¹ý°ú ÁöÁö¼¼Æ÷ÀÇ Á¾·ù¿Í mitomycin C ó¸® ½Ã°£ÀÌ ³»¼¼Æ÷±« colony Çü¼º·ü¿¡ ¹ÌÄ¡´Â ¿µÇâÀ» °üÂûÇϱâ À§ÇØ ½ÃÇàµÇ¾ú´Ù.
¿¬±¸¹æ¹ý: ÀϹÝÀûÀÎ ¸é¿ªÀýÁ¦¼ú, ÁÖ»ç¹Ù´ÃÀ» ÀÌ¿ëÇÑ ºÎºÐ ¿µ¾ç¸·¼¼Æ÷ Àý°³¹ý, Æ÷¹è±â ¹è¾Æ °ø¹è ¾ç¹ýÀ¸·Î ³»¼¼Æ÷±«¸¦ ºÐ¸®ÇÑ ÈÄ, »ó¾÷ÀûÀ¸·Î ±¸ÀÔÀÌ °¡´ÉÇÑ STO ¶Ç´Â Á÷Á¢ Á¦Á¶ÇÑ »ýÁã ¹è¾Æ¼¶ À¯¾Æ¼¼Æ÷ (pMEF)¸¦ ÁöÁö¼¼Æ÷·Î ÀÌ¿ëÇÏ¿© ¹è¾çÇÏ¿´´Ù. ¶ÇÇÑ, mitomycin C¸¦ 1, 2, 3½Ã°£ µ¿¾È ó¸®ÇÑ °¢°¢ÀÇ ÁöÁö¼¼Æ÷¿¡¼ 7ÀÏ µ¿¾È ¹è¾çÇÑ ÈÄ, ³»¼¼Æ÷±« colony Çü¼º·üÀ» »ìÆ캸¾Ò´Ù.
°á °ú: STO ÁöÁö¼¼Æ÷¿¡¼´Â ºÎºÐ ¿µ¾ç¸·¼¼Æ÷ Àý°³¹ýÀ» »ç¿ëÇÑ °æ¿ì (52%)°¡ ¸é¿ªÀýÁ¦¼ú (12%) À̳ª Æ÷¹è±â ¹è¾Æ °ø¹è¾ç¹ý (16%)À» »ç¿ëÇÑ °æ¿ìº¸´Ù ³»¼¼Æ÷±« colony Çü¼º·üÀÌ À¯ÀÇÇÏ°Ô ³ô¾Ò´Ù (p<0.05). pMEF ÁöÁö¼¼Æ÷¿¡¼ÀÇ Çü¼º·üÀº ºÎºÐ ¿µ¾ç¸·¼¼Æ÷ Àý°³¹ýÀ» »ç¿ëÇÑ °æ¿ì (88%)¿Í Æ÷¹è±â ¹è¾Æ °ø¹è¾ç¹ý (82%)À» »ç¿ëÇÑ °æ¿ì°¡ ¸é¿ªÀýÁ¦¼ú (16%)À» »ç¿ëÇÑ °æ¿ìº¸´Ù ³ô¾Ò´Ù (p<0.05). STO¿Í pMEF ¸ðµÎ¿¡¼, 2½Ã°£ mitomycin C 󸮱º (52%, 88%)ÀÌ 1½Ã°£ 󸮱º (9%, 42%)°ú 3½Ã°£ 󸮱º (18%, 76%)º¸´Ù ³ôÀº ³»¼¼Æ÷±« colony Çü¼º·üÀ» º¸¿©ÁÖ¾ú´Ù (p<0.05).
°á ·Ð: ÀÌ»óÀÇ °á°ú´Â ºÎºÐ ¿µ¾ç¸·¼¼Æ÷ Àý°³¹ýÀÌ »ýÁã Æ÷¹è±â ¹è¾Æ·ÎºÎÅÍ ³»¼¼Æ÷±«¸¦ ºÐ¸®ÇÏ´Â °¡Àå È¿°úÀûÀÎ ¹æ¹ýÀ̸ç, °¡Àå ÀûÀýÇÑ mitomycin C ó¸® ½Ã°£Àº 2½Ã°£À̶ó´Â °ÍÀ» º¸¿©ÁØ´Ù. ±×·¯³ª ÀÌ¿Í °°Àº ºÎºÐ ¿µ¾ç¸·¼¼Æ÷ Àý°³¹ýÀÇ È¿¿ë¼ºÀ» º¸´Ù ¸íÈ®ÇÏ°Ô È®ÀÎÇϱâ À§Çؼ´Â ºÐ¸®ÇÑ ³»¼¼Æ÷±«¸¦ °è´ë¹è¾çÇÏ¿© Áٱ⼼Æ÷ÁַμÀÇ Æ¯¼ºÀ» È®ÀÎÇÏ´Â ½ÇÇèÀÌ Ãß°¡ÀûÀ¸·Î ÇÊ¿äÇÒ°ÍÀ¸·Î »ý°¢µÈ´Ù.
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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³»¼¼Æ÷±« colony;ÁöÁö¼¼Æ÷;³»¼¼Æ÷±« ºÐ¸®¹æ¹ý;Mitomycin C
ICM colony;Feeder layer;Isolation method;Mitomycin C
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