An in vitro system for the study of matrix metalloproteases during decidualization in the mouse.
An in vitro system for the study of matrix metalloproteases during decidualization in the mouse.
复制标题
用于研究小鼠蜕膜化过程中基质金属蛋白酶的体外系统。
DOI：
10.1139/o96-096
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Babiarz,B
中科院分区：
文献类型：
--
作者：
Romagnano,L;Afonso,S;Babiarz,B
蜕膜化导致细胞外基质的重塑，I型胶原蛋白的丢失和基底膜基质成分的出现。我们开发了一种体外测定系统来研究小鼠蜕膜化过程中的基质金属蛋白酶。从妊娠7.5天的小鼠分离的子宫基质或蜕膜在三维I型胶原基质（Vitrogen）上生长。从这些文化的条件培养基的明胶酶谱显示在65，62，和59 kDa的62 kDa占主导地位的明胶酶A的加工形式的组成性分泌。 明胶酶A的表达模式相似，从组织裂解物的蜕膜化子宫从5.5至7.5天的发展。在Vitrogen上培养的细胞，而不是在塑料或基质包被的培养皿上培养的细胞，能够将酶原加工成体内观察到的59 kDa形式。 只有基质细胞培养的涂层的胶原蛋白I型显示相同的增加59 kDa酶原。 在Vitrogen上生长的蜕膜细胞附着，然后迁移成聚集体，这些聚集体最终穿透凝胶并作为分化的蜕膜在下面的塑料上扩散。这些初步结果表明，在体外测定系统可以用来研究的作用，金属蛋白酶在基质重塑过程中的蜕膜化。关键词：基质金属蛋白酶，子宫基质，蜕膜，明胶酶A，Vitrogen。
Decidualization results in the remodeling of the extracellular matrix with the loss of collagen type I and the appearance of basement membrane matrix components. We have developed an in vitro assay system to study matrix metalloproteases during mouse decidualization. Uterine stroma, or decidua isolated from day 7.5 pregnant mice, were grown on a three-dimensional collagen type I matrix (Vitrogen). Gelatin zymography of conditioned media from these cultures showed constitutive secretion of processed forms of gelatinase A at 65, 62, and 59 kDa with 62 kDa predominating. Similar patterns of gelatinase A expression were obtained from tissue lysates of decidualizing uteri from days 5.5 to 7.5 of development. Cells cultured on Vitrogen, but not on plastic or matrix-coated dishes, were able to process the proenzyme to the 59 kDa form as observed in vivo. Only stroma cells cultured on a coating of collagen type I displayed the same increase in the 59 kDa zymogen. Decidua cells grown on Vitrogen attached and then migrated into aggregates that eventually penetrated the gel and spread as differentiated decidua on the underlying plastic. These preliminary results suggested that the in vitro assay system can be used to study the role of metalloproteases in matrix remodeling during decidualization.Key words: matrix metalloproteases, uterine stroma, decidua, gelatinase A, Vitrogen.