Osteopetrotic mouse stroma with thrombopoietin, c-kit ligand, and flk-2 ligand supports long-term mobilized CD34+ hematopoiesis in vitro Journal Article
| Authors: | Feugier, P.; Li, N.; Jo, D. Y.; Shieh, J.H.; Mackenzie, K. L.; Lesesve, J. F.; Latger-Cannard, V.; Bensoussan, D.; Crystal, R. G.; Rafii, S.; Stoltz, J. F.; Moore, M. A. S. |
| Article Title: | Osteopetrotic mouse stroma with thrombopoietin, c-kit ligand, and flk-2 ligand supports long-term mobilized CD34+ hematopoiesis in vitro |
| 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. © Mary Ann Liebert, Inc. |
| Keywords: | controlled study; human cell; nonhuman; flow cytometry; animal cell; mouse; animals; mice; cd34 antigen; stem cell factor; cell line; membrane proteins; in vitro study; animalia; genetic vectors; cytokine; chemotaxis; peripheral blood stem cell; hematopoietic cell; murinae; cell migration; cell culture techniques; stem cells; hematopoiesis; adenovirus vector; flt3 ligand; adjuvants, immunologic; macrophage; stroma cell; stromal cells; cell adhesion; granulocyte colony stimulating factor; antigens, cd34; megakaryocyte; cd14 antigen; granulocyte; colony stimulating factor 1; colony forming unit gm; adenoviridae; antigens, cd14; coculture; coculture techniques; thrombopoietin; stromal cell derived factor 1; granulocyte-macrophage colony-stimulating factor; burst forming unit e; colony forming unit gemm; albers schoenberg disease; macrophage colony-stimulating factor; osteopetrosis |
| Journal Title: | Stem Cells and Development |
| Volume: | 14 |
| Issue: | 5 |
| ISSN: | 1547-3287 |
| Publisher: | Mary Ann Liebert, Inc |
| Date Published: | 2005-10-01 |
| Start Page: | 505 |
| End Page: | 516 |
| Language: | English |
| DOI: | 10.1089/scd.2005.14.505 |
| PUBMED: | 16305336 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 11" - "Export Date: 24 October 2012" - "CODEN: SCDTA" - "Source: Scopus" |
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