Kono Ken, Takada Nozomi, Yasuda Satoshi, Sawada Rumi, Niimi Shingo, Matsuyama Akifumi, Sato Yoji
Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan.
Division of Cellular & Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan; Research on Disease Bioresources, Platform of Therapeutics for Rare Disease and Health Policy, National Institute of Biomedical Innovation, Kobe International Business Center Rm#602, 5-5-2 Minatojima-Minami-Machi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
Biologicals. 2015 Mar;43(2):146-9. doi: 10.1016/j.biologicals.2014.11.007. Epub 2014 Dec 16.
The analysis of in vitro cell senescence/growth after serial passaging can be one of ways to show the absence of immortalized cells, which are frequently tumorigenic, in human cell-processed therapeutic products (hCTPs). However, the performance of the cell growth analysis for detection of the immortalized cellular impurities has never been evaluated. In the present study, we examined the growth rates of human mesenchymal stem cells (hMSCs, passage 5 (P = 5)) contaminated with various doses of HeLa cells, and compared with that of hMSCs alone. The growth rates of the contaminated hMSCs were comparable to that of hMSCs alone at P = 5, but significantly increased at P = 6 (0.1% and 0.01% HeLa) or P = 7 (0.001% HeLa) within 30 days. These findings suggest that the cell growth analysis is a simple and sensitive method to detect immortalized cellular impurities in hCTPs derived from human somatic cells.
连续传代后体外细胞衰老/生长分析可作为一种方法，用于证明人细胞处理的治疗产品（hCTP）中不存在通常具有致瘤性的永生化细胞。然而，用于检测永生化细胞杂质的细胞生长分析性能从未得到评估。在本研究中，我们检测了被不同剂量HeLa细胞污染的人间充质干细胞（hMSC，第5代（P = 5））的生长速率，并与单独的hMSC进行比较。在P = 5时，受污染的hMSC的生长速率与单独的hMSC相当，但在30天内，在P = 6（0.1%和0.01% HeLa）或P = 7（0.001% HeLa）时显著增加。这些发现表明，细胞生长分析是一种简单且灵敏的方法，可用于检测源自人体体细胞的hCTP中的永生化细胞杂质。
