Adhesive and degradative properties of human placental cytotrophoblast cells in vitro.
Adhesive and degradative properties of human placental cytotrophoblast cells in vitro.
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DOI：
10.1083/jcb.109.2.891
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发表时间：
1989-08
期刊：
影响因子：
--
通讯作者：
Damsky CH
中科院分区：
文献类型：
--
作者：
Fisher SJ;Cui TY;Zhang L;Hartman L;Grahl K;Zhang GY;Tarpey J;Damsky CH
人类胎儿的发育依赖于胚胎迅速进入母体循环。人类胎盘胎儿部分的滋养细胞通过短暂地表现出某些类似肿瘤的特性，解决了这个问题。因此，在妊娠早期，胎儿细胞滋养细胞侵入子宫及其动脉网络。这一过程在怀孕第12周达到顶峰，随后迅速下降，这表明细胞滋养层细胞的高度特化、侵袭性行为受到密切调节。由于对实际机制知之甚少，我们开发了一种从不同胎龄胎盘中分离细胞滋养细胞的方法，以研究其体外粘附和侵袭特性。从妊娠1、2、3个月的人胎盘中分离的细胞滋养细胞被镀在由PF HR9畸胎瘤细胞系产生的基底膜样细胞外基质上。所有孕期的细胞都表达钙依赖性细胞粘附分子cam 120/80 （e -钙粘蛋白），在胎盘中，这种分子对细胞滋养细胞是特异性的。然而，只有在前三个月，细胞滋养层细胞才会降解它们所培养的基质，在基底膜底物上留下很大的间隙，并将低分子质量的3h标记的基质成分释放到培养基中。将妊娠中期或晚期的细胞滋养细胞、妊娠早期的人胎盘成纤维细胞或人绒毛膜癌细胞系BeWo和JAR在放射性标记基质上培养时，没有观察到类似的降解活性。为了开始了解这种降解行为的生化基础，底物凝胶技术被用于分析妊娠早期、中期和晚期细胞滋养细胞表达的细胞相关和分泌的蛋白酶活性。几种明胶降解蛋白酶在妊娠早期侵袭性细胞滋养层细胞中有独特的表达，这些活性均可被金属蛋白酶抑制剂所抑制。在妊娠中期早期，细胞滋养层细胞侵袭性在体内迅速减弱，此时细胞滋养层细胞的蛋白酶模式与足月的非侵袭性细胞相同。这些结果是第一个表明特化的、暂时调节的金属蛋白酶参与滋养细胞侵袭子宫的证据。由于来自妊娠早期和妊娠后期胎盘的细胞滋养细胞在体内可维持数天暂时调节的降解行为，因此本文描述的短期细胞滋养细胞生长培养系统应有助于研究人类胎盘的一些早期事件
Human fetal development depends on the embryo rapidly gaining access to the maternal circulation. The trophoblast cells that form the fetal portion of the human placenta have solved this problem by transiently exhibiting certain tumor-like properties. Thus, during early pregnancy fetal cytotrophoblast cells invade the uterus and its arterial network. This process peaks during the twelfth week of pregnancy and declines rapidly thereafter, suggesting that the highly specialized, invasive behavior of the cytotrophoblast cells is closely regulated. Since little is known about the actual mechanisms involved, we developed an isolation procedure for cytotrophoblasts from placentas of different gestational ages to study their adhesive and invasive properties in vitro. Cytotrophoblasts isolated from first, second, and third trimester human placentas were plated on the basement membrane-like extracellular matrix produced by the PF HR9 teratocarcinoma cell line. Cells from all trimesters expressed the calcium-dependent cell adhesion molecule cell-CAM 120/80 (E-cadherin) which, in the placenta, is specific for cytotrophoblasts. However, only the first trimester cytotrophoblast cells degraded the matrices on which they were cultured, leaving large gaps in the basement membrane substrates and releasing low molecular mass 3H-labeled matrix components into the medium. No similar degradative activity was observed when second or third trimester cytotrophoblast cells, first trimester human placental fibroblasts, or the human choriocarcinoma cell lines BeWo and JAR were cultured on radiolabeled matrices. To begin to understand the biochemical basis of this degradative behavior, the substrate gel technique was used to analyze the cell-associated and secreted proteinase activities expressed by early, mid, and late gestation cytotrophoblasts. Several gelatin-degrading proteinases were uniquely expressed by early gestation, invasive cytotrophoblasts, and all these activities could be abolished by inhibitors of metalloproteinases. By early second trimester, the time when cytotrophoblast invasion rapidly diminishes in vivo, the proteinase pattern of the cytotrophoblasts was identical to that of term, noninvasive cells. These results are the first evidence suggesting that specialized, temporally regulated metalloproteinases are involved in trophoblast invasion of the uterus. Since the cytotrophoblasts from first trimester and later gestation placentas maintain for several days the temporally regulated degradative behavior displayed in vivo, the short-term cytotrophoblast outgrowth culture system described here should be useful in studying some of the early events in human placen