Combinatorial stem cell mobilization in animal models

TY - JOUR

T1 - Combinatorial stem cell mobilization in animal models

AU - Pitchford, Simon C

AU - Rankin, Sara M

PY - 2012

Y1 - 2012

N2 - It has long been recognized that single therapies, such as G-CSF, have a limited capacity to mobilize hematopoietic progenitor cells from the bone marrow. As a consequence in ∼20% of patients insufficient numbers of HPCs are mobilized to perform a bone marrow transplant. Recent studies have shown synergistic mobilization of HPCs when G-CSF pretreatment is combined with acute administration of a CXCR4 antagonist suggesting that combinatorial therapies may have therapeutic potential. In addition to HPCs, endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) reside in the bone marrow. These progenitor cells contribute to tissue regeneration and there is currently much interest in identifying the factors and mechanisms that regulate their mobilization. We describe a methodology for an in situ perfusion system of the mouse hind limb that permits direct quantification of stem and progenitor cell egress from the bone marrow. Progenitor cells are quantified by colony forming assays and immunohistochemistry. A strength of the methodology described is the ability to simultaneously quantify the mobilization of HPCs, EPCs and MSCs. Using this system we have shown that it is possible to achieve differential mobilization of these stem cell subsets using discrete combination therapies. Identification of such novel pharmacological regimens that stimulate the selective mobilization of EPCs and MSCs might be exploited in the future for tissue regeneration.

AB - It has long been recognized that single therapies, such as G-CSF, have a limited capacity to mobilize hematopoietic progenitor cells from the bone marrow. As a consequence in ∼20% of patients insufficient numbers of HPCs are mobilized to perform a bone marrow transplant. Recent studies have shown synergistic mobilization of HPCs when G-CSF pretreatment is combined with acute administration of a CXCR4 antagonist suggesting that combinatorial therapies may have therapeutic potential. In addition to HPCs, endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) reside in the bone marrow. These progenitor cells contribute to tissue regeneration and there is currently much interest in identifying the factors and mechanisms that regulate their mobilization. We describe a methodology for an in situ perfusion system of the mouse hind limb that permits direct quantification of stem and progenitor cell egress from the bone marrow. Progenitor cells are quantified by colony forming assays and immunohistochemistry. A strength of the methodology described is the ability to simultaneously quantify the mobilization of HPCs, EPCs and MSCs. Using this system we have shown that it is possible to achieve differential mobilization of these stem cell subsets using discrete combination therapies. Identification of such novel pharmacological regimens that stimulate the selective mobilization of EPCs and MSCs might be exploited in the future for tissue regeneration.

KW - Animals

KW - Cell Culture Techniques

KW - Colony-Forming Units Assay

KW - Endothelial Cells

KW - Granulocyte Colony-Stimulating Factor

KW - Hematopoietic Stem Cell Mobilization

KW - Hematopoietic Stem Cells

KW - Heterocyclic Compounds

KW - Mesenchymal Stromal Cells

KW - Mice

KW - Models, Animal

U2 - 10.1007/978-1-61779-943-3_12

DO - 10.1007/978-1-61779-943-3_12

M3 - Article

C2 - 22890929

SN - 1940-6029

VL - 904

SP - 139

EP - 154

JO - Methods in molecular biology (Clifton, N.J.)

JF - Methods in molecular biology (Clifton, N.J.)

IS - N/A

M1 - N/A

ER -