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[Abstract] [Full Text] [PDF]
An in vitro method to study the effects of hematopoietic regulators during immune and blood cell development Nitixa Patel1, Marianne Castillo1 and Pranela Rameshwar1* 1 Department of Medicine, UMDNJ-New Jersey Medical School, Newark, NJ. UMDNJ-New Jersey Medical School, MSB, Rm. E-579 185 South Orange Ave, Newark, NJ 07103, USA. New Jersey.
Phone: +(973) 972 0625
Fax: +(973) 972 8854
Email: rameshwa@umdnj.edu.
Biol. Proced. Online 2007;9:56-64. doi:10.1251/bpo133 Indexing terms: cell culture techniques; hematopoiesis; cytokines; hematopoietic stem cells. Abbreviations: BM, Bone marrow; CLP, common lymphoid progenitors; CMP, common myeloid progenitors; HSCs, hematopoietic stem cells; MSCs, mesenchymal stem cells; NK-A, neurokinin-A; NK1, neurokinin-1; NK2, neurokinin-2; SDF-1α, Stromal cell-derived factor 1α; SP, Substance P. |
In adults, hematopoiesis occurs in bone marrow (BM) through a complex process with differentiation of hematopoietic stem cells (HSCs) to immune and blood cells. Human HSCs and their progenitors express CD34. Methods on hematopoietic regulation are presented to show the effects of the chemokine, stromal-derived growth factor (SDF)-1α and the neuropeptide, substance P (SP). SDF-1α production in BM stroma causes interactions with HSCs, thereby retaining the HSCs in regions close to the endosteum, at low oxygen. Small changes in SDF-1α levels stimulate HSC functions through direct and indirect mechanisms. The indirect method occurs by SP production, which stimulates CD34+ cells, supported by ligand-binding studies, long-term culture-initiating cell assays for HSC functions, and clonogenic assays for myeloid progenitors. These methods can be applied to study other hematopoietic regulators. |