四川大学学报（医学版）
四川大學學報（醫學版）
사천대학학보（의학판）
JOURNAL OF SICHUAN UNIVERSITY(MEDICAL SCIENCE EDITION)
2010年
2期
261-264
,共4页
卵巢组织%玻璃化冷冻%卵泡%形态%增殖细胞核抗原
卵巢組織%玻璃化冷凍%卵泡%形態%增殖細胞覈抗原
란소조직%파리화냉동%란포%형태%증식세포핵항원
Ovarian tissue%Vitrification%Follicle%Morphology%PCNA
目的 探讨玻璃化冷冻对成人卵巢组织中原始卵泡与初级卵泡的形态学及细胞增殖状态的影响.方法 采用液氮直接覆盖玻璃化冷冻(direct cover vitrification,DCV)方法冻存成人卵巢组织块,液氮保存2周后复苏组织.观察和比较冻融前后人卵巢组织中原始卵泡与初级卵泡组织形态学改变;免疫组化染色测定经体外培养48 h后新鲜和冻融卵巢组织中增殖细胞核抗原(PCNA)的表达情况.结果 冻融前后人卵巢组织内原始卵泡和初级卵泡分布差异无统计学意义(P>0.05);新鲜卵巢组织中,形态异常的原始卵泡比例与初级卵泡比例差异无统计学意义(P>0.05),冻融卵巢组织中,形态异常的原始卵泡比例低于形态异常的初级卵泡比例(P<0.05);新鲜组、新鲜及冻融后培养组均有PCNA的阳性表达,PCNA 阳性表达可见于中间型卵泡和初级卵泡的卵母细胞和颗粒细胞、卵巢组织间质细胞.结论 利用DCV方法对人卵巢组织进行冻存能够保存原始卵泡和初级卵泡,冻融过程对原始卵泡和初级卵泡形态结构都可能造成一定程度的损伤,冻融后组织内卵泡有体外生长发育的潜力.
目的 探討玻璃化冷凍對成人卵巢組織中原始卵泡與初級卵泡的形態學及細胞增殖狀態的影響.方法 採用液氮直接覆蓋玻璃化冷凍(direct cover vitrification,DCV)方法凍存成人卵巢組織塊,液氮保存2週後複囌組織.觀察和比較凍融前後人卵巢組織中原始卵泡與初級卵泡組織形態學改變;免疫組化染色測定經體外培養48 h後新鮮和凍融卵巢組織中增殖細胞覈抗原(PCNA)的錶達情況.結果 凍融前後人卵巢組織內原始卵泡和初級卵泡分佈差異無統計學意義(P>0.05);新鮮卵巢組織中,形態異常的原始卵泡比例與初級卵泡比例差異無統計學意義(P>0.05),凍融卵巢組織中,形態異常的原始卵泡比例低于形態異常的初級卵泡比例(P<0.05);新鮮組、新鮮及凍融後培養組均有PCNA的暘性錶達,PCNA 暘性錶達可見于中間型卵泡和初級卵泡的卵母細胞和顆粒細胞、卵巢組織間質細胞.結論 利用DCV方法對人卵巢組織進行凍存能夠保存原始卵泡和初級卵泡,凍融過程對原始卵泡和初級卵泡形態結構都可能造成一定程度的損傷,凍融後組織內卵泡有體外生長髮育的潛力.
목적 탐토파리화냉동대성인란소조직중원시란포여초급란포적형태학급세포증식상태적영향.방법 채용액담직접복개파리화냉동(direct cover vitrification,DCV)방법동존성인란소조직괴,액담보존2주후복소조직.관찰화비교동융전후인란소조직중원시란포여초급란포조직형태학개변;면역조화염색측정경체외배양48 h후신선화동융란소조직중증식세포핵항원(PCNA)적표체정황.결과 동융전후인란소조직내원시란포화초급란포분포차이무통계학의의(P>0.05);신선란소조직중,형태이상적원시란포비례여초급란포비례차이무통계학의의(P>0.05),동융란소조직중,형태이상적원시란포비례저우형태이상적초급란포비례(P<0.05);신선조、신선급동융후배양조균유PCNA적양성표체,PCNA 양성표체가견우중간형란포화초급란포적란모세포화과립세포、란소조직간질세포.결론 이용DCV방법대인란소조직진행동존능구보존원시란포화초급란포,동융과정대원시란포화초급란포형태결구도가능조성일정정도적손상,동융후조직내란포유체외생장발육적잠력.
Objective To compare the morphology and proliferation of primordial and primary follicles in fresh and vitrificated human ovarian tissues. Methods Human ovarian tissues were cryopreserved by direct cover vitrification (DCV) for 2 weeks. The morphology of the primordial and primary follicles from the frozen-thawed tissues was compared with those from the fresh tissues. Both fresh and cryopreservation tissues were cultured for 48 hours before the tissues were embedded in paraffin block for immunohistochemical staining for PCNA. Results The distribution of primordial and primary follicles in the fresh ovarian tissues was not different from that in the frozen tissues. The cryopreserved tissues had less abnormal morphology in primordial follicles than in primary follicles, but no difference was found between the cryopreserved tissues and fresh tissues. Positive staining on PCNA expression in granulsa cells and oocyte of transitional follicles and primary follicles as well as stromal cells were found in fresh, fresh cultured and cryopreserved cultured ovarian tissues. The fresh tissues had less positive staining on PCNA in the follicle than in the fresh cultured and cryopresered cultured tissues. Conclusion Cryopreserved human ovarian tissues by DCV can maintain partial primordial and primary follicles. Follicles in cryopreserved ovarian tissues can initiate development in vitro culture.
