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Toxicologic Pathology
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Articles

Near-optimal Conditions for the In Vitro Culture of Hemopoietic Progenitor Cells in Bone Marrow from the Rat

Gemma Molyneux1,2, Sian Rizzo1, John Turton2, Parvinder Phul1,2 and Frances Gibson1

1 Division of Cellular and Molecular Medicine, Section for Cellular and Molecular Pathology, St George’s, University of London, United Kingdom
2 Centre for Toxicology, Department of Pharmacology, School of Pharmacy, University of London, United Kingdom

Correspondence: Dr. G. Molyneux, Breakthrough Breast Cancer Centre, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK; e-mail:gemma.molyneux{at}icr.ac.uk.

In vitro techniques for the culture of hemopoietic stem cells and committed hemopoietic progenitor cells in rat bone marrow have not been adequately described in the literature. In the present investigations, and using commercially available hemopoietic cytokines and growth factors, the conditions required to perform long-term bone marrow culture (LTBMC) using rat femoral bone marrow were studied, in conjunction with the colony-forming unit cell assay (CFU-C), to quantify the number of progenitor cells. CFU-C production by LTBMCs, set up using Iscove’s modified Dulbecco’s medium supplemented with fetal calf serum and horse serum, ceased after week 2 of culture. However, the duration of CFU-C production was significantly increased by supplementing LTBMCs with the cytokine recombinant mouse stem cell factor or recombinant rat stem cell factor.

Key Words: long-term bone marrow culture • rat • hemopoietic progenitor cells • colony-forming unit cell assay • mouse stem cell factor • in vitro culture

Abbreviations: BFU-E, burst-forming unit erythroid • BM, bone marrow • CFU-C, colony-forming unit cell • CFU-GEM, colony-forming units granulocyte–macrophage and erythroid elements • CFU-GM, colony-forming units granulocyte–macrophage • FCS, fetal calf serum • HS, horse serum • IMDM, Iscove’s modified Dulbecco’s medium • LTBMC, long-term bone marrow culture • rhEpo, recombinant human erythropoietin • rhG-CSF, recombinant human granulocyte colony–stimulating factor • rhIL-6, recombinant human interleukin 6 • rmIL-3, recombinant mouse interleukin 3 • rmSCF, recombinant mouse stem cell factor • rrSCF, recombinant rat stem cell factor • SCF, stem cell factor

Toxicologic Pathology, Vol. 37, No. 2, 170-174 (2009)
DOI: 10.1177/0192623308328133
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