江苏农业学报
江囌農業學報
강소농업학보
JIANGSU JOURNAL OF AGRICULTURAL SCIENCES
2015年
4期
834-839
,共6页
叶幼荣%张璐%孙杰%桑永明%赵宗胜
葉幼榮%張璐%孫傑%桑永明%趙宗勝
협유영%장로%손걸%상영명%조종성
蛋鸡%子宫内膜%上皮细胞%基质细胞%性激素%体外增殖
蛋鷄%子宮內膜%上皮細胞%基質細胞%性激素%體外增殖
단계%자궁내막%상피세포%기질세포%성격소%체외증식
laying hen%endometrium%epithelial cell%stromal cell%sex hormone%proliferation in vitro
为探讨蛋鸡子宫内膜细胞的形态学特点、生长规律,本试验采用Ⅰ型胶原酶消化的方法获取蛋鸡子宫内膜上皮细胞和基质细胞,采用免疫组织化学法进行细胞鉴定,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐比色( MTT)法测得细胞生长曲线,并研究不同浓度水平的性腺激素对子宫内膜上皮细胞和基质细胞体外增殖的影响。结果显示,该方法培养的上皮细胞和基质细胞分别对角蛋白质ck18单克隆抗体和波形蛋白质抗体呈阳性;蛋鸡子宫内膜上皮细胞和基质细胞在接种后第2 d开始进入对数生长期,第7 d进入平台期;孕酮(100 nmol/L)能显著促进子宫内膜上皮细胞和基质细胞体外增殖;孕酮(100 nmol/L)和10 nmol/L、100 nmol/L雌激素组合对子宫内膜上皮细胞和基质细胞体外增殖均有显著促进作用。表明本研究成功培养了蛋鸡子宫内膜上皮细胞和基质细胞,并证实这2种细胞的细胞周期均为7 d,一定浓度的孕酮和雌激素及其组合对子宫内膜上皮细胞和基质细胞的体外增殖均有促进作用。
為探討蛋鷄子宮內膜細胞的形態學特點、生長規律,本試驗採用Ⅰ型膠原酶消化的方法穫取蛋鷄子宮內膜上皮細胞和基質細胞,採用免疫組織化學法進行細胞鑒定,採用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴鹽比色( MTT)法測得細胞生長麯線,併研究不同濃度水平的性腺激素對子宮內膜上皮細胞和基質細胞體外增殖的影響。結果顯示,該方法培養的上皮細胞和基質細胞分彆對角蛋白質ck18單剋隆抗體和波形蛋白質抗體呈暘性;蛋鷄子宮內膜上皮細胞和基質細胞在接種後第2 d開始進入對數生長期,第7 d進入平檯期;孕酮(100 nmol/L)能顯著促進子宮內膜上皮細胞和基質細胞體外增殖;孕酮(100 nmol/L)和10 nmol/L、100 nmol/L雌激素組閤對子宮內膜上皮細胞和基質細胞體外增殖均有顯著促進作用。錶明本研究成功培養瞭蛋鷄子宮內膜上皮細胞和基質細胞,併證實這2種細胞的細胞週期均為7 d,一定濃度的孕酮和雌激素及其組閤對子宮內膜上皮細胞和基質細胞的體外增殖均有促進作用。
위탐토단계자궁내막세포적형태학특점、생장규률,본시험채용Ⅰ형효원매소화적방법획취단계자궁내막상피세포화기질세포,채용면역조직화학법진행세포감정,채용3-(4,5-이갑기새서-2)-2,5-이분기사담서추염비색( MTT)법측득세포생장곡선,병연구불동농도수평적성선격소대자궁내막상피세포화기질세포체외증식적영향。결과현시,해방법배양적상피세포화기질세포분별대각단백질ck18단극륭항체화파형단백질항체정양성;단계자궁내막상피세포화기질세포재접충후제2 d개시진입대수생장기,제7 d진입평태기;잉동(100 nmol/L)능현저촉진자궁내막상피세포화기질세포체외증식;잉동(100 nmol/L)화10 nmol/L、100 nmol/L자격소조합대자궁내막상피세포화기질세포체외증식균유현저촉진작용。표명본연구성공배양료단계자궁내막상피세포화기질세포,병증실저2충세포적세포주기균위7 d,일정농도적잉동화자격소급기조합대자궁내막상피세포화기질세포적체외증식균유촉진작용。
Endometrial epithelial and stromal cells were isolated and identified by type I collagenase digestion and immunohistochemistry, respectively. The growth curves were constructed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tet-razolium bromide ( MTT) assay. The effects of different concentrations of sex hormones, estrogen ( E2 ) and progesterone ( P4 ) , on the proliferation of the endometrial epithelial and stromal cells in vitro was studied. The epithelial and stromal cells were tested positive to keratin ck18 and vimentin antibody. The growth rates of chicken endometrial epithelial and stro-mal cells began to increase logarithmically on day 2, reaching the plateau phase on day 7. The endometrial cells and stromal cells treated with 100 nmol/L P4 proliferated at a significantly higher rate than the control. However, the endometrial epithelial cells and stromal cells treated with 100 nmol/L or 10 nmol/L E2 plus 100 nmol/L P4 proliferated at a significantly higher rate than the control.