Ex vivo expansions and transplantations of mouse bone marrow-derived hematopoietic stem/progenitor cells |
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Authors: | Email author" target="_blank">Wang?Jin-fuEmail author Wu?Yi-fan Harrintong?Jenny Ian?K?McNiece |
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Institution: | (1) College of Life Sciences, Zhejiang University, 310012 Hangzhou, China;(2) BMT Lab, University of Colorado Health Science Center, 80262 Denver, CO, USA |
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Abstract: | To examine the effects of co-culture with bone marrow mesenchymal stem cells on expansion of hematopoietic stem/progenitor
cells and the capacities of rapid neutrophil engraftment and hematopoietic reconstitution of the expanded cells, we expanded
mononuclear cells (MNCs) and CD34+/c-kit+ cells from mouse bone marrow and transplanted the expanded cells into the irradiated mice. MNCs were isolated from mouse
bone marrow and CD34+/c-kit+ cells were selected from MNCs by using MoFlo Cell Sorter. MNCs and CD34+/c-kit+ cells were co-cultured with mouse bone marrow-derived mesenchymal stem cells (MSCs) under a two-step expansion. The expanded
cells were then transplanted into sublethally irradiated BDF1 mice. Results showed that the co-culture with MSCs resulted
in expansions of median total nucleated cells, CD34+ cells, GM-CFC and HPP-CFC respectively by 10.8-, 4.8-, 65.9- and 38.8-fold for the mononuclear cell culture, and respectively
by 76.1-, 2.9-, 71.7- and 51.8-fold for the CD34+/c-kit+ cell culture. The expanded cells could rapidly engraft in the sublethally irradiated mice and reconstitute their hematopoiesis.
Co-cultures with MSCs in conjunction with two-step expansion increased expansions of total nucleated cells, GM-CFC and HPP-CFC,
which led us to conclude MSCs may create favorable environment for expansions of hematopoietic stem/progenitor cells. The
availability of increased numbers of expanded cells by the co-culture with MSCs may result in more rapid engraftment of neutrophils
following infusion to transplant recipients.
Project supported by NIH-Blood, Heart & Lung (National Institute of Health, USA, IR 01 4L70593-01) and Zhejiang Provincial
Science Foundation (No. 011103397), China |
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Keywords: | Hematopoietic stem cells Mesenchymal stem cells Expansion Transplantation |
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