Department of Obstetrics and Gynaecology, University Hospital, K.U. Leuven, BE-3000 euven, Belgium. roland.devlieger @ uz.kuleuven.ac.be
Gynecol Obstet Invest. 2010;69(1):62-6. doi: 10.1159/000255956. Epub 2009 Nov 7.
There is paucity of data on the capacity of fetal membranes to repair surgical defects following trauma. We aimed at developing an in vitro model using monolayers of human amnion epithelial cells to study fetal membrane healing.
Term (n = 6) and preterm (n = 3) fetal membranes were collected at caesarean section. The amnion was digested twice in a trypsin solution. Amniocytes were seeded (250,000-750,000/ml) and incubated at 37 degrees C and 5% CO(2) and 21 or 5% O(2). A microsurgical injury was made centrally in the monolayers and the cultures were incubated for 48 h. Every 6 h, slides were fixed and immunohistochemical staining was performed to quantify proliferation at the site of the defect and centrally in the monolayer. The closure rate was evaluated by measuring the defect size every 6 h.
The closure rate of the defects was higher in preterm versus term cultures. Proliferation was significantly higher in the defect zone versus the peripheral zone, and also higher in the preterm group.
We describe a new model for the study of fetal membrane healing and observed gestational age-dependent repair capacity of the amnion.
关于胎膜在创伤后修复外科缺损的能力的数据很少。我们旨在通过使用单层人羊膜上皮细胞建立体外模型来研究胎膜愈合。
在剖宫产时收集足月（n=6）和早产（n=3）胎膜。羊膜在胰蛋白酶溶液中消化两次。将羊膜细胞（250,000-750,000/ml）接种并在 37°C 和 5%CO(2)及 21 或 5%O(2)下孵育。在单层的中央进行微外科损伤，孵育 48 小时。每 6 小时固定载玻片并进行免疫组织化学染色，以定量测量缺陷部位和单层中央的增殖情况。每 6 小时测量缺陷大小以评估闭合率。
与足月培养物相比，早产培养物中的缺陷闭合率更高。增殖在缺陷区域明显高于周边区域，在早产组中也更高。
我们描述了一种用于研究胎膜愈合的新模型，并观察到了羊膜随胎龄修复能力的差异。
