Price J M
Am J Anat. 1979 Sep;156(1):147-57. doi: 10.1002/aja.1001560116.
Sertoli cells have been insolated from the newborn calf testis using a combination of mechanical and enzymatic disruption. Testicular fragments, previously chopped into 1-mm pieces, are digested in an enzyme mixture consisting of hyaluronidase, collagenase, trypsin and DNAse, followed by a second digestion in trypsin and DNAse. Isolation of the resulting cellular fractions by sedimentation with unit gravity produces an aliquot of Sertoli cells which is over 95% pure when examined by light and electron microscopy. Cultures of these cells grow rapidly and produce Mullerian Inhibiting Substance as evidenced by their ability to cause the involution of the Mullerian duct of the female fetal rat when co-cultured in an organ-culture assay system.
利用机械和酶解相结合的方法从新生小牛睾丸中分离出支持细胞。预先切成1毫米碎片的睾丸组织块，在由透明质酸酶、胶原酶、胰蛋白酶和脱氧核糖核酸酶组成的酶混合物中消化，然后再用胰蛋白酶和脱氧核糖核酸酶进行二次消化。通过单位重力沉降分离得到的细胞组分，得到的支持细胞等分试样在光学显微镜和电子显微镜检查时纯度超过95%。这些细胞培养物生长迅速，并产生苗勒氏管抑制物质，在器官培养测定系统中共培养时，它们能够使雌性胎鼠的苗勒氏管退化，这证明了这一点。
