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Anin vitro method to study the effects of hematopoietic regulators during immune and blood cell development

Abstract

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.

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

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Correspondence to Pranela Rameshwar Ph.D..

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Patel, N., Castillo, M. & Rameshwar, P. Anin vitro method to study the effects of hematopoietic regulators during immune and blood cell development. Biol. Proced. Online 9, 56–64 (2007). https://doi.org/10.1251/bpo133

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  • DOI: https://doi.org/10.1251/bpo133

Indexing terms

  • cell culture techniques
  • hematopoiesis
  • cytokines
  • hematopoietic stem cells