Premium
Immortalization without neoplastic transformation of human mesenchymal stem cells by transduction with HPV16 E6/E7 genes
Author(s) -
Hung ShihChieh,
Yang DenMei,
Chang ChingFang,
Lin RueyJen,
Wang JihShiuan,
LowTone Ho Larry,
Yang Wen K.
Publication year - 2004
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.20126
Subject(s) - mesenchymal stem cell , biology , stem cell , neoplastic transformation , cell culture , microbiology and biotechnology , population , cell growth , transduction (biophysics) , cell , cellular differentiation , immunology , cancer research , genetics , gene , carcinogenesis , medicine , biochemistry , environmental health
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 CD34 dim CD38 + 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.