Analysis of inter-species cross-contamination in 160 non-human cell lines / 基础医学与临床

Objective To analyze interspecies cross-contamination of 160 non-human cell lines.Methods One hundred and sixty common non-human cell lines were collected and their species were identified by PCR.For the suspicious cells,chromosome analysis was further used to confirm their species.Results Six in 160 non-human cell lines were cross-contaminated.A rat cell line was mixed by a human cell line,and 5 were totally cross-con-taminated,and were indentified as wrong species.Conclusions Species identification is an indispensable part of cell quality control.Each cell line should undergo a full QA(Quality Assurance)assessment before it is used for research.

Effect of JAK/STAT pathway activation on high glucose-induced transdifferentiation in renal proximal tubular epithelial cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the effect of JAK/STAT signaling pathway activation on the transdifferentiation and secretion of transforming growth factor-beta1 (TGF-beta1) induced by high glucose in renal proximal tubular epithelial cells.</p><p><b>METHODS</b>Human kidney cells (HKC) were cultured and then divided into four groups: low glucose (LG) group, high glucose (HG) group, high mannitol (LG + M) group, and HG + AG490 group. Immunoprecipitation and Western blot analysis were used to determine the expression of tryosine phosphorylated Janus kinase 2 ( p-JAK2). The protein expressions of STAT1, STAT3, p-STAT1, and p-STAT3 and the expressions of alpha-SMA and E-Cadherin were observed by Western blot. The contents of TGF-B1, fibronectin and type I collagen in the supernatants of the cultured HKC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-beta1 mRNA was measured by reverse transcription and polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Compared with LG group, the expressions of JAK2, p-STAT1, p-STAT3, and TGF-beta1, mRNA were significantly increased in HG group from 6 to 72 hours. Meanwhile, the contents of TGF-beta1 and collagen I in the supernatants and the expression of alpha-SMA increased and the expression of E-Cadherin decreased. The expressions of JAK2, p-STAT1, p-STAT3, and TGF-beta mRNA as well as the levels of TGF-beta1 and collagen I in the supernatant s in HG + AG490 group were significantly lower than in the HG group. The expressions of alpha-SMA and E-Cadherin were also decreased in HG + AG490 group.</p><p><b>CONCLUSION</b>Activation of JAK/STAT signaling pathway may be involved in the high glucose-induced transdifferentiation and overproduction of TGF-beta1, and ECM proteins in HKCs.</p>