Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

doi:10.1016/j.bbrc.2006.05.169

Biochemical and Biophysical Research Communications

Volume 347, Issue 1, 18 August 2006, Pages 12-21

https://doi.org/10.1016/j.bbrc.2006.05.169 Get rights and content

Abstract

Low oxygen tension is a potent differentiation inducer of numerous cell types and an effective stimulus of many gene expressions. Here, we described that under 8% O₂, bone marrow stromal cells (MSCs) exhibited proliferative and morphologic changes. The level of differentiated antigen H-2Dd and the number of G₂/S/M phase cells increased evidently under 8% O₂ condition. Also, the proportion of wide, flattened, and epithelial-like cells (which were alkaline phosphatase staining positive) in MSCs increased significantly. When cultured in adipogenic medium, there was a 5- to 6-fold increase in the number of lipid droplets under hypoxic conditions compared with that in normoxic culture. We also demonstrated the existence of MSC differentiation under hypoxic conditions by electron microscopy. Expression of Oct4 was inhibited under 8% O₂ condition, but after adipocyte differentiation in normoxic culture and hypoxia-mimicking agents cobalt chloride (CoCl₂) and deferoxamine mesylate (DFX) treatments, Oct4 was still expressed in MSCs. These results indicate hypoxia accelerates MSC differentiation and hypoxia and hypoxia-mimicking agents exert different effects on MSC differentiation.

Section snippets

Materials and methods

Animals. Eight- to 12-week-old male and female Balb/c mice were used as bone marrow (BM) donors. All mice were purchased from Laboratory Animal Center (Beijing University School of Medicine). All animal handle and experimental procedures were approved by the Animal Care and Use Committee of the Chinese Academy of Medical Sciences.

Isolation of MSCs from bone marrow and culture conditions. BM was collected from the tibial and femoral diaphysis. The ends of bones were cut, and the marrow was

Proliferative response of MSCs to low oxygen and CoCl₂

It has been reported that hypoxia could drive proliferation of human pulmonary artery fibroblasts and the mechanism was also discussed [17]. However, the effect of low oxygen on MSC differentiation and proliferation has not been evaluated. We examined the protein levels of HIF-1α under normoxic conditions, 8% low oxygen conditions and normoxic conditions with 100 μM CoCl₂. The protein level of HIF-1α was undetectable under normoxic condition, but significantly increased when MSCs were cultured

Discussion

MSCs-based therapy is of potential value in tissue replacement and regeneration, and has been studied in clinical trials in autologous and allogeneic settings [22]. Recent studies have shown that the proportion of osteogenesis cells under hypoxic conditions was greater than under normoxic conditions in vitro and in vivo [16]. Our results in the study showed that the marrow stromal cells under our culture contained very primitive cells which were Oct4-positive and have adipocyte differentiation

Acknowledgments

This work was supported by the “863 Projects” of Ministry of Science and Technology of PR China (No. 2002AA205061); from China Medical Board of New York, Inc: Stem Cell Biology, Engineering (Grant #01-748); from National Natural Science Foundation of China (No. 30125018); from National Key Project for Basic Research of China (No. 001CB5099); R.C. Zhao is a Cheung Kong Scholar in PR China.

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¹: These authors contributed equally to this work.

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Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

Abstract

Section snippets

Materials and methods

Proliferative response of MSCs to low oxygen and CoCl2

Discussion

Acknowledgments

Exp. Hematol.

Blood

Blood

Blood

Blood

Blood

Cell

J. Biol. Chem.

Prockop marrow stromal cells as stem cells for nonhematopoietic tissues

Science

Pluripotency of mesenchymal stem cells derived from adult marrow

Nature

Establishment and properties of fetal dermis-derived mesenchymal stem cell lines: plasticity in vitro and hematopoietic protection in vivo

Bone Marrow Transplant

In vivo and in vitro differentiation of myocytes from human bone marrow-derived multipotent progenitor cells

Exp. Hematol.

Mesenchymal stem cell injection induces cardiac nerve sprouting and increased tenascin expression in a Swine model of myocardial infarction

J. Cardiovasc. Electrophysiol.

Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth

Proc. Natl. Acad. Sci. USA

Targeting gene expression to hypoxic tumor cells

Nat. Med.

Simultaneous activation of several second messengers in hypoxia-induced hyperpermeability of brain derived endothelial cells

J. Cell. Physiol.

Cobalt chloride and low oxygen tension trigger differentiation of acute myeloid leukemic cells: possible mediation of hypoxia-inducible factor-1alpha

Leukemia

Hypoxia modifies proliferation and differentiation of CD34(+) CML cells

Stem Cells

Proliferative response of MSCs to low oxygen and CoCl₂