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Beneficial reciprocal effects of bone marrow stromal cells and Schwann cells from adult rats in a dynamic co?culture system in vitro without intercellular contact.

ABSTRACT: In order to examine how implanted bone marrow stromal cells (BMSCs) encourage peripheral nerve regeneration, the present study investigated the interaction of BMSCs and Schwann cells (SCs) using an indirect in vitro co?culture model. SCs and BMSCs were obtained from adult Sprague?Dawley rats. The passaged BMSCs were CD29? and CD44?positive but CD45?negative and were co?cultured with the primary SCs using a Millicell system, which allows BMSCs and SCs to grow in the same culture medium but without direct contact. Expression of the typical SC markers S?100 and glial fibrillary acidic protein (GFAP) of the treated BMSCs as well as the proliferation capacity of the co?cultured SCs was evaluated by immunocytochemical staining on the 3rd and 5th day of co?culture. Immunocytochemical staining showed that >75% of the BMSCs in the indirect co?culture model were GFAP? and S?100?positive on the 3rd and 5th day after co?culture, as opposed to <5% of the BMSCs in the control group. On the 3rd day after co?culture, only a few co?cultured BMSCs showed the typical SC?like morphology, while most BMSCs still kept their native appearance. By contrast, on the 5th day after co?culture, almost all of the co?cultured BMSCs appeared with the typical SC?like morphology. Furthermore, 70.71% of the SCs in the indirect co?culture model were S?100?positive on the 5th day of co?culture, as opposed to >30.43% of the SCs in the control group. These results indicated that BMSCs may interact synergistically with SCs with regard to promoting peripheral nerve regeneration.

SUBMITTER: Zhou LN

PROVIDER: S-EPMC4581791 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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Beneficial reciprocal effects of bone marrow stromal cells and Schwann cells from adult rats in a dynamic co‑culture system in vitro without intercellular contact.

Zhou Li-Na LN Cui Xiao-Jun XJ Su Kai-Xin KX Wang Xiao-Hong XH Guo Jin-Hua JH

Molecular medicine reports 20150702 4

In order to examine how implanted bone marrow stromal cells (BMSCs) encourage peripheral nerve regeneration, the present study investigated the interaction of BMSCs and Schwann cells (SCs) using an indirect in vitro co‑culture model. SCs and BMSCs were obtained from adult Sprague‑Dawley rats. The passaged BMSCs were CD29‑ and CD44‑positive but CD45‑negative and were co‑cultured with the primary SCs using a Millicell system, which allows BMSCs and SCs to grow in the same culture medium but withou ...[more]

PMID: 26133460

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Project description:BackgroundAngiogenesis is a critical part of the endogenous repair process in brain injury and disease, and requires at least two sequential steps. First, angiogenic sprouting of endothelial cells occurs, which entails the initial proliferation of endothelial cells and remodeling of the surrounding extracellular matrix. Second, vessel stabilization is necessary to prevent vascular regression, which relies on vascular smooth muscle recruitment to surround the young vessels. Marrow stromal cells (MSCs) have been shown to promote revascularization after hindlimb ischemia, cardiac ischemia, and stroke. SB623 cells are derived from marrow stromal cells by transfection with a Notch1 intracellular domain (NICD)-expressing plasmid and are known to elicit functional improvement in experimental stroke. These cells are currently used in human clinical testing for treatment of chronic stroke. In the current study, the angiogenic property of SB623 cells was investigated using cell-based assays.MethodsAngiogenic paracrine factors secreted by SB623 cells and the parental MSCs were identified using the Qantibody Human Angiogenesis Array. To measure the angiogenic activity of conditioned medium from SB623 cells and MSCs, endothelial tube formation in the human umbilical vein endothelial cell (HUVEC) assay and endothelial cell sprouting and branching in the rodent aortic ring assay were quantified. To validate the angiogenic contribution of VEGF in conditioned medium, endothelial cells and aortic rings were treated with SU5416, which inhibits VEGFR2 at low dose.ResultsConditioned medium from SB623 cells promoted survival and proliferation of endothelial cells under serum-deprived conditions and supports HUVEC vascular tube formation. In a rodent aortic ring assay, there was enhanced endothelial sprouting and branching in response to SB623-derived conditioned medium. SU5416 treatment partially reversed the effect of conditioned medium on endothelial cell survival and proliferation while completely abrogate HUVEC tube formation and endothelial cell sprouting and branching in aortic ring assays.ConclusionsThese data indicate that SB623 cell-secreted angiogenic factors promoted several aspects of angiogenesis, which likely contribute to promoting recovery in the injured brain.

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Beneficial reciprocal effects of bone marrow stromal cells and Schwann cells from adult rats in a dynamic co?culture system in vitro without intercellular contact.

Publications

Beneficial reciprocal effects of bone marrow stromal cells and Schwann cells from adult rats in a dynamic co‑culture system in vitro without intercellular contact.

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