| 학회지 검색 | HOME > 논문 및 학회지 > 학회지 검색 |
대한생식의학회지 제27권 제3호 2010년
생쥐 난소에서 Preantral Follicle의 단순 분리법
대구대학교 축산학과1, 경북대학교병원 산부인과학교실2
김주환1, 박기상1,2, 송해범1, 전상식2,
A Simple Isolating Method of Preantral Follicles from Mouse Ovaries
Ju Hwan Kim1, Kee Sang Park1,2, Hai Bum Song1, Sang Sik Chun2
1Department of Animal1, Science, Taegu University 2Department of Obstetrics and Gynecology, Kyungpook National University Hospital
Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1)enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at 37℃ and 5% CO2 incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 IU/ml and DNAse 20 IU/ml. After 20 min, follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when p value was less than 0.05. Results: In handing time, mincing or scraping method (28±3.42 min or 16±1.58 min) were significantly (p<0.00001) shorter than enzymatical method (72±1.69 min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method (49.8±3.91) than in mincing or scraping method (25.3±2.33 or 20.5±1.75). Isolated follicles in ≤90 um were significantly (p<0.005) higher in enzymatical method (15±1.71) than in mincing or scraping method (7.8±0.98 or 8.1±1.31). In 91~130 um, isolated follicles were significantly (p<0.0005) higher in enzymatical method (33±3.27) than in mincing or scraping method (163±1.82 or 10.7±1.38). In ≥131 um, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ≤90 um was highest in scraping method (39.6% vs. enzymatical Method: 30.1%, p<0.05; mincing Method: 30.9%, p=0.11719, NS). Rate of follicles in 91~130 um was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ≥131 um was highest in scraping method (8.3% vs. enzymatical or scraping Method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handing time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91~130 um was highest in all methods.
키워드 : Preantral follicles from mouse ovaries, Enzymatical method, Mincing method, Scraping method
교신저자 : .
전문 파일 : 
