Devore-Carter D, Morway P F, Weiss E B
Division of Respiratory Diseases, Saint Vincent Hospital, Worcester, Massachusetts 01604.
Cell Tissue Res. 1988 Feb;251(2):325-31. doi: 10.1007/BF00215840.
A simple 30-min enzyme digestion procedure has been used to release guinea-pig tracheal smooth muscle cells that retain differentiated function in long-term subculture. Primary cell cultures initially consist of numerous epithelial colonies and 70-1000 morphologically differentiated smooth muscle cells per 600 mg (wet weight) tracheal tissue depending on the age of the animal. Both cell types proliferate to form a confluent monolayer within 5-17 days. Pure subcultures of tracheal smooth muscle cells are obtained by limited trypsin digestion of the primary culture. Eighty percent of these subcultured smooth muscle cells retain the ability to contract in response to histamine (10(-6) M) and to form reaggregates even after 20 or more passages. Examination of these cells by electron microscopy reveals both biosynthetic and contractile components of smooth muscle. Analysis of this dual phenotype may provide valuable information about the regulation of tracheal smooth muscle cell growth and differentiation.
一种简单的30分钟酶消化程序已被用于释放豚鼠气管平滑肌细胞，这些细胞在长期传代培养中仍保留分化功能。原代细胞培养最初由大量上皮集落和每600毫克（湿重）气管组织中70 - 1000个形态学上分化的平滑肌细胞组成，这取决于动物的年龄。两种细胞类型都会增殖，在5 - 17天内形成汇合的单层。通过对原代培养物进行有限的胰蛋白酶消化可获得气管平滑肌细胞的纯传代培养物。这些传代培养的平滑肌细胞中，80%即使在传代20次或更多次后仍保留对组胺（10⁻⁶ M）作出收缩反应并形成再聚集物的能力。通过电子显微镜检查这些细胞可发现平滑肌的生物合成和收缩成分。对这种双重表型的分析可能会提供有关气管平滑肌细胞生长和分化调节的有价值信息。
