We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
An improved method for isolating Schwann cells from postnatal rat sciatic nerves.
- Authors
Yujun Wei; Jianli Zhou; Zhenghuan Zheng; Aijun Wang; Qiang Ao; Yandao Gong; Xiufang Zhang
- Abstract
The major difficulty in Schwann cell (SC) purification is contamination by fibroblasts, which usually become the predominant cell type during SC enrichment in vitro. Current reported measures are mainly limited by either high cost or complicated procedures with low cell yields or purity. Our objectives have been to develop an efficient, easily applicable, rapid method to obtain highly purified SC from the sciatic nerve of newborn rats. The method involves two rounds of purification to eliminate fibroblasts with the novel combined use of cytosine-B-arabinoside hydrochloride (Ara-C) action and differential cell detachment. Cultured cells were first treated with Ara-C for 24 h. The medium was replaced with the growth medium containing 20 ng/ml human heregulin1-β1 extracellular domain (HRG1-β1 ECD). After another 48 h in culture, the cells were treated with 0.05% trypsin, following which SCs, but not fibroblasts, were easily detached from the dishes. The advantage of this method is that the two steps can eliminate the fibroblasts complementarily. Ara-C eliminates most of the fibroblasts growing among SCs, whereas the differential cell detachment technique removes the remainder growing under or interacting with the SC layer. A purity of more than 99% SCs has been obtained, as confirmed by cell morphology and immunostaining. The purified SCs have a spindle-shaped, bipolar, and sometimes tripolar morphology, align in fascicles, and express S-100. The whole procedure takes about 10 days from primary culture to the purified SCs growing to confluence (only half the time reported previously). This protocol provides an alternative method for investigating peripheral nerve regeneration and potentially could be used to produce enough SCs to construct artificial nerve scaffolds in vitro.
- Subjects
FIBROBLASTS; CELLS; NERVOUS system regeneration; MURIDAE; DIGESTIVE enzymes
- Publication
Cell & Tissue Research, 2009, Vol 337, Issue 3, p361
- ISSN
0302-766X
- Publication type
Article
- DOI
10.1007/s00441-009-0836-4