人肝细胞生长因子杂交瘤细胞株的建立  

Establishment or Hybridoma Cell Strain Secreting Human Hepatocyte Growth Factor

作　　者：韩伟国[1] 杨伟珠 钱宇平[1] 吕伟中[2] 张利能[3] 查锡良[3] 

机构地区：[1]镇江医学院生化教研室,镇江212001 [2]镇江市第一人民医院,镇江212001 [3]上海医科大学生化教研室,上海200032

出　　处：《中国生物制品学杂志》2000年第2期72-74,共3页Chinese Journal of Biologicals

基　　金：江苏省教委资助项目(95197)

摘　　要：目的　建立能分泌人肝细胞生长因子（ｈＨＧＦ）的杂交瘤细胞株。方法　从４～５月龄的人胎肝中分 离贮脂细胞，与次黄嘌呤鸟嘌呤磷酸核糖转移酶（ＨＧＰＲＴ）缺陷型人肝癌细胞株（ＳＭＭＣ－７７２１）融合成杂交瘤细胞，用 超滤膜对培养上清液初步分离，并用Ｈ３－ＴｄＲ掺入法测定各组分中ｈＨＧＦ的生物活性。结果　培养上清液中相对分 子质量 １０ ０００～ ３０ ０００的组分ｈＨＧＦ活性显著增高，对小鼠肝细胞增殖有明显的促进作用，并存在量效依赖关系，而 相对分子质量小于１０ ０００或大于 ３０ ０００的组分与对照相比 ｈＨＧＦ活性无明显改变。结论 ｈＨＧＦ杂交瘤细胞的建 立可作为获得ｈＨＧＦ的一条新途径。Objective To establish a hybridoma cell strain secreting human hepatocyte growth factor (hHGF) .Methods Fat-storing cells were isolated from 4-5 months old fetal liver and fused with a hypo- thantin-guanine phospho-ribosyl transferase(HGPRT)deficient unman hepatoma cell strain(SMMC-7721). The culture supernatant was fractionated by ultrafiltration and the biological activity of hHGF in the fractions was determined by H3-TdR penetration method. Results The biological activity of hHGF in the fraction with a molecular weight of 10 000-30 000 was significantly improved. However, compared with control, no significant changes were observed in the activities of fractions with the moleculr weights of less than 10 000 or more than 30 000.The hHGF secreted by the strain showed a dose-dependent promoting effect on the proliferation of murine hepatocytes. Conclusion The establishment of hygridoma cell strain secreting hHGF might be a new route for obtaining hHGF.

关 键 词：肝细胞生长因子 次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT) 肝癌细胞株(SMMG-7721) 杂交瘤细胞 

分 类 号：Q813.2[生物学—生物工程] Q513.203