Joshi M S
Biol Reprod. 1985 Aug;33(1):187-200. doi: 10.1095/biolreprod33.1.187.
A simple and rapid method has been described to isolate the epithelial cells of cauda epididymidis of adult bull. Perfusion of the lumen of the cauda epididymidis with 1 mg/ml collagenase in calcium- and magnesium-free Hank's balanced salt solution and incubation of the tissue at 37 degrees C for 90 min releases the principal and basal cells into the lumen. Several individual epithelial cells and cell aggregates without the contamination of stromal or smooth muscle cells can be flushed out at the end of incubation. The isolated epithelial cells, suspended in Dulbecco's medium with 10% horse serum, attach to plastic dishes within 3 h after seeding the cells and proliferate to form a monolayer in approximately 8-12 days. The electron microscopic study and immunostaining of the cultured epithelial cells indicate that the cultured cells are principal cells. The basal cells of the intact cauda epididymidis of bull show within their cytoplasm the presence of varying amounts of "lipid-like" material often closely associated with whorls of membrane.
已描述了一种简单快速的方法来分离成年公牛附睾尾部的上皮细胞。用不含钙和镁的汉克平衡盐溶液配制的1mg/ml胶原酶灌注附睾尾部管腔，并将组织在37℃孵育90分钟，可使主细胞和基底细胞释放到管腔中。孵育结束时，可以冲出几个未受基质或平滑肌细胞污染的单个上皮细胞和细胞聚集体。分离的上皮细胞悬浮于含10%马血清的杜尔贝科培养基中，接种后3小时内附着于塑料培养皿，并在约8-12天内增殖形成单层。对培养的上皮细胞进行电子显微镜研究和免疫染色表明，培养的细胞是主细胞。公牛完整附睾尾部的基底细胞在其细胞质内显示存在不同数量的“类脂质”物质，这些物质通常与膜的螺旋紧密相关。
