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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 LN, Cui XJ, Su KX, Wang XH, Guo JH - Mol Med Rep (2015)

Bottom Line: 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.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.These results indicated that BMSCs may interact synergistically with SCs with regard to promoting peripheral nerve regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China.

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.

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BMSCs as well as SCs grown in the upper HA membranes developed well at the 2nd and 5th day of co-culture, as shown by hematoxylin and eosin staining (scale bar, 200 mm). (A) BMSCs were still present an had formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture. (B) BMSCs gradually appeared with an SC-like morphology on the 5th day of co-culture. (C) SCs developed the typical spindle-like shape with a round nucleus in the centre of the cell body and two or three slender processes, and grew in clusters on the 5th day of co-culture. BMSC, bone marrow stromal cell; SC, Schwann cell.
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f5-mmr-12-04-4931: BMSCs as well as SCs grown in the upper HA membranes developed well at the 2nd and 5th day of co-culture, as shown by hematoxylin and eosin staining (scale bar, 200 mm). (A) BMSCs were still present an had formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture. (B) BMSCs gradually appeared with an SC-like morphology on the 5th day of co-culture. (C) SCs developed the typical spindle-like shape with a round nucleus in the centre of the cell body and two or three slender processes, and grew in clusters on the 5th day of co-culture. BMSC, bone marrow stromal cell; SC, Schwann cell.

Mentions: The well-grown BMSCs in the upper HA membranes formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture, as shown by hematoxylin and eosin (HE) staining (Fig. 5A), while the BMSCs in the upper HA membranes exhibited a typical SC-like morphology with spindle-shaped or triangular soma and two or three slender processes on the 5th day of co-culture (Fig. 5B). Similarly, the primary SCs grown in the upper HA membranes developed the typical spindle-shaped soma with two or three slender processes. On the 5th day of co-culture, the SCs grown in the upper HA membranes grew in clusters and their densities were markedly higher than those of the fibroblasts, as shown by HE staining (Fig. 5C).


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 LN, Cui XJ, Su KX, Wang XH, Guo JH - Mol Med Rep (2015)

BMSCs as well as SCs grown in the upper HA membranes developed well at the 2nd and 5th day of co-culture, as shown by hematoxylin and eosin staining (scale bar, 200 mm). (A) BMSCs were still present an had formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture. (B) BMSCs gradually appeared with an SC-like morphology on the 5th day of co-culture. (C) SCs developed the typical spindle-like shape with a round nucleus in the centre of the cell body and two or three slender processes, and grew in clusters on the 5th day of co-culture. BMSC, bone marrow stromal cell; SC, Schwann cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581791&req=5

f5-mmr-12-04-4931: BMSCs as well as SCs grown in the upper HA membranes developed well at the 2nd and 5th day of co-culture, as shown by hematoxylin and eosin staining (scale bar, 200 mm). (A) BMSCs were still present an had formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture. (B) BMSCs gradually appeared with an SC-like morphology on the 5th day of co-culture. (C) SCs developed the typical spindle-like shape with a round nucleus in the centre of the cell body and two or three slender processes, and grew in clusters on the 5th day of co-culture. BMSC, bone marrow stromal cell; SC, Schwann cell.
Mentions: The well-grown BMSCs in the upper HA membranes formed an adherent monolayer with a mesenchymal morphology on the 2nd day of co-culture, as shown by hematoxylin and eosin (HE) staining (Fig. 5A), while the BMSCs in the upper HA membranes exhibited a typical SC-like morphology with spindle-shaped or triangular soma and two or three slender processes on the 5th day of co-culture (Fig. 5B). Similarly, the primary SCs grown in the upper HA membranes developed the typical spindle-shaped soma with two or three slender processes. On the 5th day of co-culture, the SCs grown in the upper HA membranes grew in clusters and their densities were markedly higher than those of the fibroblasts, as shown by HE staining (Fig. 5C).

Bottom Line: 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.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.These results indicated that BMSCs may interact synergistically with SCs with regard to promoting peripheral nerve regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China.

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.

Show MeSH
Related in: MedlinePlus