A novel method for obtaining highly enriched Schwann cell populations from mature monkey nerves based on in vitro pre‑degeneration

Wang,Gangyang; Ma,Zhengwen; Cao,Lingling; Yan,Guofeng; Wang,Yang; Jin,Yuqing; Shen,Hua; Zhang,Yiping; Xu,Xiaoming; Chen,Xuejin; Shen,Zunli

doi:10.3892/mmr.2017.7427

A novel method for obtaining highly enriched Schwann cell populations from mature monkey nerves based on in vitro pre‑degeneration

Authors:
- Gangyang Wang
- Zhengwen Ma
- Lingling Cao
- Guofeng Yan
- Yang Wang
- Yuqing Jin
- Hua Shen
- Yiping Zhang
- Xiaoming Xu
- Xuejin Chen
- Zunli Shen
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Affiliations: Department of Plastic and Reconstructive Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China, Department of Laboratory Animal Science, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China, Department of Rehabilitation Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China, Norton Neuroscience Institute, Norton Healthcare, Louisville, KY 40202, USA, Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery, Indiana University School of Medicine, Indianapolis, IN 46202, USA
Published online on: September 5, 2017 https://doi.org/10.3892/mmr.2017.7427
Pages: 6600-6607
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Schwann cells (SCs) are indispensable for cell therapy and tissue engineering of the peripheral nervous system. Easy access to activated, highly proliferative SCs is necessary for clinical applications. The present study developed a fast, efficient method for obtaining highly purified SCs from the peripheral nerve of a mature Rhesus monkey. The common peroneal nerves of 4‑year‑old Rhesus monkeys were harvested and subjected to in vitro pre‑degeneration in a modified SC culture medium (SCCM). The nerve pieces were subsequently treated enzymatically to dissociate the cells and then cultured for 2 days in SCCM. Cultured cells were treated with purification medium containing Ara‑C to assist in restricting the overgrowth of fibroblast‑like cells, for 24 h. After another 24‑h cultivation period, the cells were subsequently treated with a multiplex collagenase, which enabled SC detachment over fibroblast detachment, and thereby facilitated SC isolation. Finally, SCs were cultured in SCCM. The cell yield was determined by cell counting following enzyme digestion and SC purity was determined from the percentage of SCs with respect to the total number of cells. Following purification, 96.3±3.9% of cells were identified as SCs. In vitro pre‑degeneration in the presence of basic‑fibroblast growth factor, heregulin β1 and forskolin maximized the purity and yield of SCs that could be obtained from monkey peroneal nerves. The present study identified a novel technique that can efficiently isolate and purify SCs from mature monkey nerves based on in vitro pre‑degeneration.

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