Hasebe-Takada Nozomi, Kono Ken, Yasuda Satoshi, Sawada Rumi, Matsuyama Akifumi, Sato Yoji
Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Tokyo, Japan.
Platform of Therapeutics for Rare Disease, National Institutes of Biomedical Innovation, Health and Nutrition, Amagasaki, Japan.
Regen Ther. 2016 Sep 9;5:49-54. doi: 10.1016/j.reth.2016.06.005. eCollection 2016 Dec.
In human cell-processed therapeutic products (hCTPs) for clinical application, tumorigenic cellular impurities in the manufacturing process are a major concern. Because cellular immortalization is one of the prerequisite steps in tumorigenesis, we tested whether cell growth analysis can be employed to check for immortalized (and potentially tumorigenic) cellular impurities in hCTPs. We monitored the growth of human bone marrow-derived mesenchymal stem cells (BMSCs) mixed with HeLa cells at a ratio of 1/10 or more and compared their growth rates with that of BMSCs alone. The cell growth analysis detected a significant increase in the growth rate of the BMSCs spiked with 0.0001% HeLa within 30 days at a probability of 47%. When human adipose-derived stem cells (ADSCs) were spiked with ASC52telo cells, a human telomerase reverse transcriptase (hTERT)-immortalized adipose-derived mesenchymal stem cell line, at a ratio of 0.001% or more, their growth rates were significantly increased within 15 passages, compared with that of ADSCs alone. These results indicate that cell growth analysis for the detection of immortalized cellular impurities in human somatic stem cells is simple and can be useful for the quality assessment of hCTPs in the manufacturing process.
在用于临床应用的人细胞处理治疗产品（hCTP）中，制造过程中的致瘤性细胞杂质是一个主要问题。由于细胞永生化是肿瘤发生的前提步骤之一，我们测试了细胞生长分析是否可用于检测hCTP中永生化（以及潜在致瘤性）细胞杂质。我们监测了以1/10或更高比例与HeLa细胞混合的人骨髓间充质干细胞（BMSC）的生长情况，并将其生长速率与单独的BMSC进行比较。细胞生长分析检测到，掺入0.0001% HeLa的BMSC在30天内生长速率显著增加，概率为47%。当人脂肪来源干细胞（ADSC）以0.001%或更高比例掺入ASC52telo细胞（一种人端粒酶逆转录酶（hTERT）永生化脂肪来源间充质干细胞系）时，与单独的ADSC相比，其在15代内生长速率显著增加。这些结果表明，用于检测人成体干细胞中永生化细胞杂质的细胞生长分析方法简单，可用于hCTP制造过程中的质量评估。
