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Osteopetrotic Mouse Stroma With Thrombopoietin, C-Kit Ligand, and flk-2 Ligand Supports Long-Term Mobilized CD34+ Hematopoiesis in Vitro

Osteopetrotic Mouse Stroma With Thrombopoietin, C-Kit Ligand, and flk-2 Ligand Supports Long-Term Mobilized CD34+ Hematopoiesis in Vitro

Pierre Feugier, Na Li, Deog-Yeon Jo, Jae Hung Shieh, Karen L MacKenzie, J F Lesesve, V Latger-Cannard, D Bensoussan, Ronald G Crystal, Shahin Rafii, J F Stoltz, Malcolm A S Moore

Stem Cells Dev. 2005 Oct;14(5):505-16.

PMID: 16305336

Abstract:

OP-9 cells are stromal cells derived from macrophage colony-stimulating factor (M-CSF)-deficient osteopetrotic mice. To evaluate the OP-9 capability to sustain long-term hematopoiesis, we reported the expansion of granulocyte colony-stimulating factor (G-CSF)-mobilized human peripheral blood (PB) CD34(+) cells in co-culture with murine OP-9 and MS-5 stromal cells, either transfected with various combinations of adenovectors (Ad) expressing c-kit ligand (KL) (either soluble or transmembrane form), thrombopoietin (TPO), flt-3/flk2 ligand (FL), and granulocyte-macrophage (GM)-CSF or with weekly addition of these cytokines. Expression of TPO as well as association of TPO, FL, and KL increased progenitor cell and week-6 cobblestone area forming cell (CAFC) production in all stromal co-cultures. Similar progenitor expansion was obtained by weekly addition of soluble cytokine. Five weeks of co-culture with OP9 and TPO, FL + KL resulted in the greatest expansion of progenitor cells and week-6 CAFC as measured by secondary assay on MS-5. In contrast to MS-5 and TPO or TPO + FL + KL cultures where hematopoiesis declined by week 4, progenitor as well as week-6 CAFC expansion continued for over 3 months in TPO + FL + KL OP9 cocultures. This was associated with decrease of CD14(+) macrophage production. The addition of human macrophage (M)-CSF or CD14(+) cells to the co-culture decrease progenitor and stem cell expansion; however, murine M-CSF to OP-9 co-cultures did not decrease progenitor expansion. High levels of stromal-derived factor-1 (SDF-1) production by MS-5 and low or absent production by OP-9 may account for stem cell adhesion and CAFC formation in the former cultures and the predominance of stem and progenitor cells in the nonadherent fraction in the latter cultures.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR4248578	Flt-3/Flk-2 ligand human			Price

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