Osteoprogenitor cells as targets for ex vivo gene transfer

J. E. Onyia, D. Clapp, H. Long, J. M. Hock

Research output: Contribution to journalArticle

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Abstract

We transduced osteoprogenitor cells with recombinant retrovirus and analyzed proviral integration patterns into chromosomal DNA to detect for the first time the clonal and cellular fate of osteoprogenitor-derived progeny cells. Metaphyseal bone cells and diaphyseal stromal cells were isolated from the distal femurs of young rats, transduced with the vM5neolacZ recombinant retrovirus, and selected in the neomycin analog, G418. Following surgical marrow ablation of a femur in one leg of mature rats, retroviral-transduced metaphyseal or diaphyseal cells were injected into the ablated site. These rats were killed 5-6 days later. Metaphyseal and diaphyseal cells were isolated from distal femurs, selected in G418, and stained for β- galactosidase (β-gal+). The number and clonal origin of transduced progenitor cells were determined. High numbers of β-galactosidase colonies with an osteoblast phenotype were obtained following metaphyseal transplants and detected in 100% of metaphyseal and none of diaphyseal specimens. In contrast, β-galactosidase colonies derived from diaphyseal transplants were detected in 50% of specimens in both the metaphysis and diaphysis, and the absolute number of progenitor cell colonies was 60-fold less than metaphyseal transplants. Provirus was only detected in the ablated bones and not in the contralateral bone or other tissues. Proviral integration fragment analysis showed a single integration site for recovered metaphyseal cell clones, consistent with their origination from a common single progenitor. This is one of the first demonstrations of successful transplantation of clonal osteoprogenitors to their site of origin in bone. It may be possible to use these cells to target genes to bone for therapeutic use in skeletal and hematopoietic diseases.

Original languageEnglish
Pages (from-to)20-30
Number of pages11
JournalJournal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
Volume13
Issue number1
DOIs
StatePublished - Jan 1998

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Galactosidases
Bone and Bones
Femur
Genes
Retroviridae
Transplants
Stem Cells
Proviruses
Diaphyses
Neomycin
Therapeutic Uses
Stromal Cells
Osteoblasts
Leg
Clone Cells
Transplantation
Bone Marrow
Phenotype
DNA
antibiotic G 418

ASJC Scopus subject areas

  • Surgery

Cite this

Osteoprogenitor cells as targets for ex vivo gene transfer. / Onyia, J. E.; Clapp, D.; Long, H.; Hock, J. M.

In: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 13, No. 1, 01.1998, p. 20-30.

Research output: Contribution to journalArticle

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