Madeja Z, Klein A
Institute of Molecular Biology, Jagellonian University, Kraków, Poland.
Folia Histochem Cytobiol. 1992;30(3):97-101.
The autocrine growth factor(s) was isolated from serumfree conditioned medium of rat sarcoma (XC) cells. Autocrine activity was enriched by ultrafiltration using Amicon YM 10 membrane, extraction with 1 M acetic acid and partially purified (650-fold) by chromatography on Bio-Gel P-100 and P-60. The final recovery of the autocrine factor(s) was 4 micrograms from 1800 ml of the conditioned medium (a yield of 6%). The factor(s) with molecular weight 6-10 kDa was heat and acid stable but inactivated by trypsin and dithiothreitol. It stimulated anchorage-dependent (but not anchorage-independent) growth of XC cells as well as untransformed, established lines of rat (NRK) and mouse (3T3) cells. The results obtained may suggest that autocrine factor(s) produced by XC cells can be one of EGF-like or/and insulin-like growth factors.
自分泌生长因子是从大鼠肉瘤(XC)细胞的无血清条件培养基中分离出来的。使用Amicon YM 10膜进行超滤、用1 M乙酸萃取并通过在Bio-Gel P-100和P-60上进行色谱法对自分泌活性进行富集并部分纯化(650倍)。从1800 ml条件培养基中最终回收的自分泌因子为4微克(产率为6%)。分子量为6-10 kDa的因子对热和酸稳定,但被胰蛋白酶和二硫苏糖醇灭活。它刺激了XC细胞以及未转化的大鼠(NRK)和小鼠(3T3)细胞系的贴壁依赖性(而非贴壁非依赖性)生长。所获得的结果可能表明,XC细胞产生的自分泌因子可能是表皮生长因子样或/和胰岛素样生长因子之一。