Branham W S, Lyn-Cook B D, Andrews A, McDaniel M, Sheehan D M
Division of Reproductive and Developmental Toxicology, Food and Drug Administration, Jefferson, Arkansas.
In Vitro Cell Dev Biol. 1991 Jun;27A(6):442-6. doi: 10.1007/BF02631142.
We have developed a tissue culture system using an extract of basement membrane (extracellular matrix) which promotes the in vitro growth and development of uterine luminal epithelium from the 5-day-old rat. Uterine luminal epithelium, free of stroma, was obtained as short tubes by trypsinization of uterine segments followed by mechanical separation. Epithelial segments were grown in a serum-free medium on culture dishes coated with an extracellular matrix. After 2 days, rapid cell growth resulted in monolayer cultures, which subsequently formed organoid structures similar to differentiated uterine glands present in uterine tissue taken from older rats. Electron microscopy of cultures revealed columnar cells with basally located nuclei, apical microvilli, lateral membranes with interdigitations, desmosomes, and secretory Golgi complexes, all features found in functioning uterine epithelium in vivo. This model will allow the in vitro investigation of the development of uterine epithelium-specific functions free of the influence of stromal cell factors.
我们利用基底膜(细胞外基质)提取物开发了一种组织培养系统,该系统可促进5日龄大鼠子宫腔上皮在体外的生长和发育。通过对子宫段进行胰蛋白酶消化,然后进行机械分离,获得了不含基质的子宫腔上皮短管。上皮段在涂有细胞外基质的培养皿中于无血清培养基中生长。2天后,细胞快速生长形成单层培养物,随后形成类似于从老年大鼠子宫组织中取出的分化子宫腺的类器官结构。培养物的电子显微镜检查显示,细胞为柱状,细胞核位于基部,顶端有微绒毛,侧面细胞膜有指状交错、桥粒和分泌性高尔基体复合物,这些都是体内功能性子宫上皮中发现的特征。该模型将允许在无基质细胞因子影响的情况下,对子宫上皮特异性功能的发育进行体外研究。