野生型组织型纤溶酶原激活剂(t-PA)工程细胞株生物学特性分析  

The analysis of biological characteristics for the wild-type t-PA engineering cell lines

作　　者：徐秀英[1] 陈琳[1] 欧阳应斌[1] 刘士辉[1] 朱恒奇[1] 黄培堂[1] 黄翠芬[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》1997年第1期14-16,共3页Letters in Biotechnology

摘　　要：野生型t-PA工程细胞4B3形态与亲代细胞CHO-dhfr-相似呈多角形,类似上皮细胞。在MTX加压达5×10^(-7)mol/L时,少数细胞略变瘦长,但仍显多角形,形态正常。细胞无血清培养上清经FAPA检测,亚克隆株表达水平为10~6细胞4000～5000IU/24h。细胞经体外传代3个月及冻存3个月后复苏,部分亚克隆株表达水平下降,多数仍为10~6细胞4000IU/24h,表明4B3细胞株稳定性好。裸鼠试验表明,该工程株活细胞、细胞DNA、纯化的4B3rt-PA均无致瘤性病毒,支原体检查为阴性。遗传特性分析表明,该工程细胞与其亲代细胞CHO-dhfr-细胞染色体数均为20条,均有不同程度不同类型的畸变。该工程细胞的畸变率为16％。CHO-dhfr-细胞畸变率为6％。属正常范围。结果表明4B3细胞株为高效表达的性能优良的工程细胞株。The morphology of wild-type t-PA engineering cell line 4B3 is similar to CHO-dhfr cell in polygon. When aminopterin (MTX) concentration is 5 × 10-7mol/L,a part of cell morphology change slim, but they are still in polygon. Therefore, the morphology of cell line is nomal. The expression level of free-serum culture is 4000-5000 UI/106cells/24h. The cell line is bequeathed for three months in vitro and revived after freeze three months, the greater part of cell are stability. The expression level is 4000 IU/106cells/24h. The assay of tumorigenesis shows that the cell, celular DNA and purifical products don't all form tumors in nude mice. The cell line hasn't be contaminated by virus and mycoplasma. The results of hereditary characteristics show that the cell chromosome is 20 and the cell distortion index is 16%, CHO-dhfr- cell us 6%. It is nomal in scope. Therefore, the engineering cell line is good.

关 键 词：野生型t-PA 工程细胞株 特性分析 

分 类 号：Q813.2[生物学—生物工程]