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Differentiation potential of conditionally immortalized mesenchymal progenitor cells from adult marrow of a H‐2K b ‐tsA58 transgenic mouse
Author(s) -
Dennis James E.,
Caplan Arnold I.
Publication year - 1996
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199606)167:3<523::aid-jcp16>3.0.co;2-4
Subject(s) - mesenchymal stem cell , progenitor cell , genetically modified mouse , microbiology and biotechnology , transgene , progenitor , biology , stem cell , genetics , gene
Abstract Primary cultures were initiated from marrow, spleen, and bone explants of an adult H‐2K b ‐tsA58 transgenic mouse (immortomouse). All cultures were initiated in immortalizing conditions, and an additional marrow culture was first incubated for 1 week in standard conditions and then switched to immortalizing conditions. Marrow cells immediately immortalized were designated the marrow immediate population (MIP); those immortalized after 1 week were termed the marrow delayed population (MDP). MIP and MDP cells both contained a mixture of fibroblastic or flattened cells, and the MIP cells contained an additional subpopulation of adipocytic (Oil Red‐O positive) cells. Alkaline phosphatase expression was induced by dexamethasone (10 −7 M) in MDP cells while MIP, spleen, and bone explant cells had only a low level of expression. MDP and MIP cells differentiated into bone when combined with porous calcium phosphate ceramics and implanted subcutaneously into nude mice while bone‐ and spleen‐derived cells did not. Clones were isolated from the MDP and MIP cell populations and tested for differentiated phenotypes. Some MIP‐derived clones exhibited adipocytic characteristics while MDP‐derived subclones were negative. Histologic examination of porous ceramic implanted clones showed that all of the clones had osteogenic potential. Clones exposed to either dexamethasone, human recombinant bone morphogenetic protein‐2, or horse serum plus hydrocortisone showed differences in expression of adipocytic or osteogenic markers. These immortalized cultures have retained both adipocytic and osteogenic potential even after 1 year of continuous culture, and provide a model system for clonal analysis of the developmental potential of marrow‐derived mesenchymal precursor cells. © 1996 Wiley‐Liss, Inc.