研究用肿瘤细胞系的质对量控制和要求.pptVIP

研究用肿瘤细胞系的质量控制和要求 中国医学科学院基础医学研究所 刘玉琴 Some Facts about the use of cultured cells-- Nobel prize 2001，Cell cycle 2002, apoptosis 2006, RNA Interference 2007, homologous recombination and gene targeting 2008,HPV—cervical ca, HIV--aids 2009, telomere telomerase 现代生命科学研究中培养细胞的使用越来越广泛 1969~2004：cell use increasing steadily Papers use cells increase 2~2.5X biology, pharmacology and medicine---- tumor research 不可少的研究材料、对象、工具 Use of contaminated or misidentified cells increase 10X Common problems microbiological contamination with particular respect to mycoplasma infections (incidence of ca. 25%) cross-contamination by other cell lines and misidentification or misclassification (incidence of more than 15%) acquisition of genetic and epigenetic variations due to over-passaging of cell lines leading to a genetic drift of cell cultures and to the selection of altered subclones. Where from? faulty cell culture techniques: shared reagents, repeated use of the same pipette during re-feeding operations and manipulation of multiple cultures at the same time without adequate isolation of one cell type from another. the introduction of false, contaminated or over-passaged cell lines from other sources into the cell culture laboratory (35% ) Mislabeling, lack of awareness Intentional co-cultivation during propagation of human stem or primary cells using a feeder layer derived from another species 细胞交叉污染历史回顾 1968,Apparent HeLa cell contamination of human heteroploid cell lines.Nature.217:750-751 1974，HeLa cess contaminating cultures around the world,Sciences 184:1059 1981, Contaminated cell cultures. Nature 289:227 1981,cross-contamination of cells in culture. Sciences. 212(4493):226-52 细胞交叉污染历史回顾 1999, False human hematopoietic cell lins: cross-cantaminations and misint erpretations. Leukemia.13(10):1601-7 2003, False leukmia –lymphoma cell lines :an update on over 500 cell lines. Leukemia 17, 416-426 2004,cell line cross-comtamination:how aware are…in vitro cell dev biol an