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Immortalization without neoplastic transformation of human mesenchymal stem cells by transduction with HPV16 E6/E7 genes
7 auth. S. Hung, D. Yang, Ching-Fang Chang, Ruey-Jen Lin, Jih‐Shiuan Wang, L. Low-Tone Ho, ...
hMSCs derived from bone marrow are useful as a species‐specific cell culture system for studying cell lineage differentiation and tissue remodeling. However, hMSCs usually have a short in vitro life span due to replicative senescence. We therefore u…
hMSCs derived from bone marrow are useful as a species‐specific cell culture system for studying cell lineage differentiation and tissue remodeling. However, hMSCs usually have a short in vitro life span due to replicative senescence. We therefore used a high dose of retroviral vector LXSN‐16E6E7 to transduce hMSCs of an aging donor and obtained an actively proliferating cell line, designated KP‐hMSCs, which expressed HPV16 E6/E7 mRNA. Whereas parental hMSCs ceased to grow after 30 PDs, KP‐hMSCs could be propagated beyond 100 PDs. With culture procedures to avoid selection pressure and crowded cell growth, KP‐hMSCs showed no signs of neoplastic transformation as examined by soft‐agar anchorage‐independent growth and NOD‐SCID mouse tumorigenicity assays. KP‐hMSCs gave similar cytofluorimetric profiles of 31 CD markers to those of the parental primary hMSCs, except with some morphologic changes and expansion of an originally very minor CD34dimCD38+CD50+ cell population. Upon exposure to specific stimulating conditions in vitro, KP‐hMSCs could respond and differentiate along the mesenchymal (bone, fat and cartilage) and nonmesenchymal (neuron) cell lineages. Our results indicated that hMSCs could be immortalized by transduction with HPV16 E6/E7, maintained without neoplastic transformation by careful culture procedures and thus useful for stem cell research and clinical application. © 2004 Wiley‐Liss, Inc.
Published in
International Journal of Cancer
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3
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6 | 2004 |
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