冻融人卵巢组织与体细胞共培养对卵泡生长发育和卵巢组织内分泌影响

Research on the co-culture of thawed human ovarian tissues and somatic cells

目的评价冻融人卵巢组织与人早孕蜕膜细胞共培养对卵巢组织内分泌和卵泡生长发育的影响。方法 2008年3月至8月将冻融的人卵巢组织与人早孕蜕膜单层细胞共培养。实验分四组:A组:经玻璃化冻融卵巢组织与人早孕蜕膜单层细胞共培养;B组:经程序化冻融卵巢组织与人早孕蜕膜单层细胞共培养;C组:经玻璃化冻融卵巢组织单独培养;D组:经程序化冻融卵巢组织单独培养。冻融卵巢组织在共培养体系中培养14d,培养期间隔天吸取1次培养液,测定培养液中雌二醇、孕酮含量,并比较培养前后冻融卵巢组织中总体卵泡存活率(TFSR)、卵泡生长率(GFR)。结果 A组和B组,TFSR培养前后比较差异无统计学意义(P>0.05),GFR培养前后比较差异有统计学意义(P<0.01);C组和D组,培养前后TFSR、GFR比较差异有统计学意义(P<0.01)。TFSR、GFR培养后A组和B组比较,差异无统计学意义,C组和D组比较差异亦无统计学意义。A组与C组、D组、B组与C组、D组分别比较TFSR、GFR均差异有统计学意义(P<0.01)。四组间雌二醇、孕酮水平差异有统计学意义(P<0.01)。结论两种冷冻方法冻融的卵巢组织经体外培养仍具有内分泌功能,原始卵泡在体外培养体系中有不同程度生长;冻融卵巢组织与人早孕蜕膜细胞共培养优于其单独培养。

Objective To evaluate the influences Oil the endocrine function of ovarian tissues and the development of the follicles by the co-culture system for thawed human ovary tissues and somatic cells. Methods March to August 2008 the co-culture system for human ovarian tissues with decidual monolayer cells of the human early pregnancy after vitrification were set up. There were four groups in the experiment ： group A, the thawed ovarian tissues after vitrification were co-cultured with the decidua monolayer ceils; group B： Co-culturing the thawed ovarian tissues after slow-freezing with the decidua monolayer cells; group C： Culturing the thawed ovarian tissues after vitrification alone; group D： Culturing the thawed ovarian tissues after slow-freezlng alone. The culture medium was collected every other day during in vitro culture for 14 days, and the level of estrogen and progesterone were measured and compared among the groups. After 14 days culturing,the total follicle survival rate（TFSR） and the grow tbllicle rate （GFR）among the groups were compared. Resuits There are no differences between before and after the freezing in group A （P 〉 0. 05） on grow follicle rate （TF- SR） , but the grow follicle rate （GFR） has statistical differences（P 〈0. 01 ）. The same results exist in group B. There are no statistical differences in TFSR and GFR between group A and B after culturing. In group C and D, the two indexes both have statistical differences before and after culturing, but no differences exists between group C and D after culturing. Comparing the two indexes between A and C, A and D, B and C, B and D, the differences were significant. The estrogen and progesterone level of the culture medium are significant different among the four groups after analysis repeat measurement. Conclusions The thawed ovarian tissues that be cultured in-vitro have endocrine function. The original follicles have different development in culture system. If the thawed ovarian tissues are co-euhured with the decidual monolayer cells of the human early pregnancy, the result is better than culturing alone.