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정연길,석상현,박성백,임여정,최선호,송해범 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
난자의 체외성숙 및 체외배양에는 일반적으로 동물의 혈청을 기본배양액에 5-10% 정도 첨가한 배양액을 사용하고 있다. 그러나, 혈청으로부터 바이러스, 세균, 마이코 플라즈마 등에 오염될 가능성이 있기 때문에, 본 실험에서는 완전 무혈청 배양액에서 난자의 성숙, 배발생, 세포수, 동결성을 검토하였다. 도축된 한우의 난소로부터 채취한 난자는 선별하여 TCM199+10% FBS와 IVMD 101 배양액에서 22~24시간 동안 체외성숙시킨 후, IVF 100(
석상현,정연길,임여정,박성백,송해범 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
근래, 혈청배지로 생산한 체외배양 수정란은 이식후, 낮은 수태율, 과체중의 산자 생산, 높은 유산율과 사망율 등 문제가 지적되고 있다. 그러나 무혈청 배지로 생산한 체외배양 수정란은 이러한 현상을 개선할 수 있다는 보고가 있어, 본 연구는 세포성장인자가 첨가된 완전 무혈청 배양액에서 생산된 수정란으로 2001년5월 부터 2002년 12월까지의 신선란 및 동결란으로 이식한 결과이다. 이식은 신선란 및 동결란에서 A급 수정란은 1개, B급 수정란은 2개를
글루코스와 피부르산염의 농도가 조절된 DMEM과 Vero 세포를 이용한 무혈청 적응용 배양액의 개발에 관한 연구
김주환 ( Ju Hwan Kim ),서영석 ( Young Seok Seo ),송해범 ( Hai Bun Song ),양정보 ( Jung Bo Yang ),이경은 ( Kyung En Lee ),이기환 ( Ki Hwan Lee ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.2
Objective: The purpose of this study was to examine in vitro development of early preimplantation mouse embryos in various kind of serum-free conditioned media (SF-VCM) manufactured from DMEM cultured with Vero Cells. Methods: A total of 846 two-cell mouse embryos were cultured in different kind of SF-VCM. SF-VCMs were divided into SF-VCM-10, -30 and -50 by media volume using DMEM #1 media, and divided into SF-VCM #1, #2 and #3 by controlled concentration of glucose and pyruvate (manufactured by DMEM #1: mixed three volume of DMEM-G (DMEM with glutamine without glucose and pyruvate) and one volume of DMEM-GGP (DMEM with glutamine, glucose, pyruvate), #2: mixed same volume of DMEM-G and DMEM-GGP and #3: mixed one volume of DMEM-G and three volume of DMEM-GGP, respectively). Experimental groups were mainly added 10% SSS, and 20% hFF was added to only Control group co-cultured with Vero cells. Development of embryos was observed every 24 hours. Results between different groups were analyzed using Chi-square test, and considered statistically significant when P-value was less than 0.05. Results: In vitro developmental rate by each cleavage stages of mouse embryos cultured in SF-VCMs with a various volumes were significantly (P<0.05) higher in SF-VCM-30 (morula≤: 97.2%, Blastocyst (BL)≤: 97.2%, Hatching BL≤: 82.2%) than other groups. In the rate of development on in vitro co-culture vs. a various SF-VCMs manufactured by DMEM controlled concentration of glucose and pyruvate, Group I (SF-VCM #1) was higher than other groups in each cleavage stages (morula≤: 98.1%, Blastocyst (BL)≤: 97.1%, hatching BL≤: 81.7%, respectively). Moreover, specially, in the developmental rate into the hatching blastocyst ≤after 96 hours in vitro culture, Group I (81.7%) was significantly higher than control group (67.6%, P<0.05). Conclusion: SF-VCM #1 manufactured by volume of 30 mL DMEM #1 media cultured in vitro for 48 hours in 250 mL flask was the most effective on in vitro developmental rate of mouse preimplantation embryos. Therefore, it is expected that SF-VCM #1 has application to human IVF-ET.