Isolation and characterization of osteoblast precursor cells from human bone marrow
Open Access
- 1 March 1996
- journal article
- research article
- Published by Oxford University Press (OUP) in Journal of Bone and Mineral Research
- Vol. 11 (3) , 312-324
- https://doi.org/10.1002/jbmr.5650110305
Abstract
Osteoblasts are derived from precursor cells present in low frequency in the stromal element of bone marrow. Because of the lack of a practical procedure to isolate osteoblast precursors from early cultures of plastic adherent cells from bone marrow, previous studies of marrow stromal cells have been made in confluent cultures of bone marrow when the osteoblast (OB) precursors are already differentiated. Also these studies utilized cultures containing mixed populations of cells including hematopoietic cells. Thus we have employed a negative immunoselection procedure to remove contaminating hematopoietic cells and to isolate nearly homogeneous populations of early human stromal cells derived from the plastic-adherent mononuclear marrow cells cultured in the presence of serum. By reverse transcriptase polymerase chain reaction (RT-PCR) analysis for mRNA, and by immunocytochemical study for protein, we studied the sequential expression in culture of multiple markers of the osteoblast phenotype-alkaline phosphatase, osteopontin, parathyroid hormone receptor, types I and III procollagen, and osteocalcin-as well as lipoprotein lipase (LPL), a marker of the adipocyte phenotype. At an early stage of culture (7-9 days), human OB precursors formed colonies of variable sizes that expressed low levels of mRNA and protein concentrations of OB markers, and their concentration increased on growth to a confluent monolayer (approximately 14 days). LPL mRNA was expressed at high levels in the colony stage, and its level decreased upon confluency, suggesting a loss of potential for commitment to the adipocyte lineage. Interestingly, treatment with dexamethasone at 10−8 M increased the expression for some of the osteoblast markers and for the LPL gene and was required for the deposition of mineralized matrix and for the formation of adipocytes containing cytoplasmic lipid droplets in confluent cultures. Cloned single early colonies were able to coexpress the osteoblast and adipocyte markers (as assessed by RT-PCR). Thus these immunoselected marrow stromal cells have the characteristics of authentic human osteoblast precursor cells which also are capable of differentiating into adipocytes.Keywords
Funding Information
- NIH (AG-04875)
This publication has 42 references indexed in Scilit:
- The non-osteogenic mouse pluripotent cell line, C3H10T1/2, is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic protein-2Published by Elsevier ,2005
- Expression of cell growth and bone specific genes at single cell resolution during development of bone tissue‐like organization in primary osteoblast culturesJournal of Cellular Biochemistry, 1992
- Characterization of cells with osteogenic potential from human marrowBone, 1992
- Formation of osteoblast‐like cells from human mononuclear bone marrow culturesAPMIS, 1991
- Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cellsJournal of Cellular Physiology, 1990
- Determination of numbers of osteoprogenitors present in isolated fetal rat calvaria cells in vitroDevelopmental Biology, 1989
- Wound Macrophages Express TGF-α and Other Growth Factors in Vivo: Analysis by mRNA PhenotypingScience, 1988
- Tissue specificity and developmental expression of rat osteopontinBiochemical and Biophysical Research Communications, 1987
- Expression of differentiated function by mineralizing cultures of chicken osteoblastsDevelopmental Biology, 1987
- Osteoporosis and the Replacement of Cell Populations of the Marrow by Adipose TissueClinical Orthopaedics and Related Research, 1971