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体外间接共培养雪旺细胞诱导大鼠脂肪间充质干细胞向雪旺样细胞分化。

Schwann-like cell differentiation of rat adipose-derived stem cells by indirect co-culture with Schwann cells in vitro.

机构信息

School of Life Sciences, State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing, China.

出版信息

Cell Prolif. 2010 Dec;43(6):606-16. doi: 10.1111/j.1365-2184.2010.00710.x.

Abstract

OBJECTIVES

Schwann cell (SC) transplantation is a promising therapy for peripheral nerve transaction, however, clinical use of SCs is limited due to their very limited availability. Adipose-derived stem cells (ADSCs) have been identified as an alternative source of adult stem cells in recent years. The aim of this study was to evaluate the feasibility of using ADSCs as a source of stem cells for differentiation into Schwann-like cells by an indirect co-culture approach, in vitro.

MATERIALS AND METHODS

Multilineage differentiation potential of the obtained ADSCs was assayed by testing their ability to differentiate into osteoblasts and adipocytes. The ADSCs were co-cultured with SCs to be induced into Schwann-like cells through proximity, using a Millicell system. Expression of typical SC markers S-100, GFAP and P75NTR of the treated ADSCs was determined by immunocytochemical staining, western blotting and RT-PCR. Myelination capacity of the differentiated ADSCs (dADSCs) was evaluated in dADSC/dorsal root ganglia neuron (DRGN) co-cultures.

RESULTS

The treated ADSCs adopted a spindle shaped-like morphology after co-cultured with SCs for 6 days. All results of immunocytochemical staining, western blotting and RT-PCR showed that the treated cells expressed S-100, GFAP and P75NTR, indications of differentiation. dADSCs could form Schwann-like cell myelin in co-culture with DRGNs. Undifferentiated ADSCs (uADSCs) did not form myelin compared to DRGNs cultured alone, but could produce neurite extension.

CONCLUSIONS

These results demonstrate that this indirect co-culture microenvironment could induce ADSCs to differentiate into Schwann-like cells in vitro, which may be beneficial for treatment of peripheral nerve injuries in the near future.

摘要

目的

施万细胞(SC)移植是周围神经损伤的一种很有前途的治疗方法,然而,由于其可用性非常有限,SC 的临床应用受到限制。近年来,脂肪来源干细胞(ADSCs)已被确定为成体干细胞的另一种来源。本研究旨在评估通过间接共培养方法将 ADSCs 用作分化为 Schwann 样细胞的干细胞源的可行性,体外。

材料和方法

通过检测成骨细胞和脂肪细胞分化能力来测定获得的 ADSCs 的多能分化潜能。通过使用 Millicell 系统,将 ADSCs 与 SC 共培养以通过接近诱导其分化为 Schwann 样细胞。通过免疫细胞化学染色、Western blot 和 RT-PCR 测定处理后的 ADSCs 表达典型的 SC 标志物 S-100、GFAP 和 P75NTR。在 dADSC/背根神经节神经元(DRGN)共培养物中评估分化的 ADSCs(dADSCs)的髓鞘形成能力。

结果

与 SC 共培养 6 天后,处理后的 ADSCs 采用梭形样形态。免疫细胞化学染色、Western blot 和 RT-PCR 的所有结果均表明,处理后的细胞表达 S-100、GFAP 和 P75NTR,表明分化。dADSCs 可与 DRGN 共培养形成 Schwann 样细胞髓鞘。与单独培养的 DRGN 相比,未分化的 ADSCs(uADSCs)未形成髓鞘,但可产生神经突延伸。

结论

这些结果表明,这种间接共培养微环境可以在体外诱导 ADSCs 分化为 Schwann 样细胞,这可能对治疗周围神经损伤有帮助。

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