Immortalization of human marrow stromal cells by retroviral transduction with a temperature sensitive oncogene: Identification of bipotential precursor cells capable of directed differentiation to either an osteoblast or adipocyte phenotype

A. Houghton, B. O. Oyajobi, G. A. Foster, R. G.G. Russell, B. M.J. Stringer

Research output: Contribution to journalArticle

43 Scopus citations

Abstract

The etiology of osteoporosis is multifactorial, but there is evidence from both animal and human studies that the volume of marrow adipose tissue increases when bone volume is reduced in osteoporosis. The cell-related mechanism that may account for this inverse relationship between the volume of marrow adipose tissue and bone remains to be clarified, although it is known that both adipocytes and osteoblasts are derived from stromal cell precursors in bone marrow. We report that retroviral transduction with a temperature-sensitive oncogene (SV40 large T antigen) can generate bipotential cell lines from human marrow stroma that are capable of directed differentiation, in vitro, down either an osteogenic or adipocytic lineage pathway. One such clone, designated hOP 7, expresses type α1(I) procollagen and has low alkaline phosphatase (AP) activity under basal culture conditions that is reminiscent of an osteoprogenitor cell. Exposure of hOP 7 cells to dexamethasone upregulates AP activity and enables the cells to mineralize their extracellular matrix. Also, treatment with calcitriol induces osteocalcin expression and both PTH and PGE2 induce/augment cAMP formation. Incubation with normal rabbit serum, however, causes the cells to become adipogenic as demonstrated by histological staining with Oil-red-O, expression of mRNA for the early and late adipocyte markers lipoprotein lipase and glycerol 3-phosphate dehydrogenase, respectively, and loss of type α1(I) procollagen mRNA. The generation of homogeneous populations of these cells, as confirmed by Southern blot analysis, demonstrates the capacity of a human clonal cell line to differentiate in either an osteogenic or adipogenic direction.

Original languageEnglish (US)
Pages (from-to)7-16
Number of pages10
JournalBone
Volume22
Issue number1
DOIs
StatePublished - Jan 1998
Externally publishedYes

Keywords

  • Adipocyte
  • Differentiation
  • Marrow
  • Osteoblast
  • Osteoporosis
  • SV40-T

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

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