Characterization of the cell growth analysis for detection of immortal cellular impurities in human mesenchymal stem cells

Ken Kono; Nozomi Takada; Satoshi Yasuda; Rumi Sawada; Shingo Niimi; Akifumi Matsuyama; Yoji Sato

doi:10.1016/j.biologicals.2014.11.007

Characterization of the cell growth analysis for detection of immortal cellular impurities in human mesenchymal stem cells

Biologicals. 2015 Mar;43(2):146-9. doi: 10.1016/j.biologicals.2014.11.007. Epub 2014 Dec 16.

Authors

Ken Kono¹, Nozomi Takada², Satoshi Yasuda³, Rumi Sawada⁴, Shingo Niimi⁵, Akifumi Matsuyama⁶, Yoji Sato⁷

Affiliations

¹ Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan. Electronic address: [email protected].
² 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. Electronic address: [email protected].
³ Division of Cellular & Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan; Foundation for Biomedical Research and Innovation, Hyogo, Japan. Electronic address: [email protected].
⁴ Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan. Electronic address: [email protected].
⁵ Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan. Electronic address: [email protected].
⁶ 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. Electronic address: [email protected].
⁷ Division of Cellular & Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo 158-8501, Japan; Foundation for Biomedical Research and Innovation, Hyogo, Japan; Department of Quality Assurance Science for Pharmaceuticals, Graduate School of Pharmaceutical Sciences, Nagoya City University, Aichi, Japan; Department of Cellular & Gene Therapy Products, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Department of Translational Pharmaceutical Sciences, Graduated School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan. Electronic address: [email protected].

PMID: 25523786
DOI: 10.1016/j.biologicals.2014.11.007

Abstract

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.

Keywords: Cellular therapy; Mesenchymal stem cell; Quality; Regenerative medicine; Safety; Tumorigenicity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques / methods*
HeLa Cells
Humans
Mesenchymal Stem Cells / cytology*