Extraction and purification of tumorigenic Schwann cells in vitro from neurofibromatosis type 2 tissues

Abstract

Objective To establish a simple method to extract and purify tumorigenic Schwann cells (TSCs) from neurofibromatosis type 2 (NF2) tumor tissues in vitro, in order to improve the successful rate of primary culture and obtain a large number of TSCs from NF2 patients for further study. Methods Six fresh tumor tissues from patients with NF2 were obtained from 2009 to 2010 in our hospital. TSCs were extracted after collagenase digestion, followed by cell purification through rapid digestion with low trypsin concentration, differential detachment and clone screening. Morphological changes of TSCs were observed under inverted microscope and cultured TSCs were identified immunocytochemically by S ⁃ 100 protein staining. Results TSCs were successfully cultured from 3 NF2 specimens. A large number of bipolar spindle or triangular⁃shaped cells could be seen on the third day of primary culture and 4 to 5 clones were formed after the third passage in each successfully cultured specimen. Purified TSCs expressed S ⁃ 100 protein detected by immunocytochemistry with the rate of positive cells of above 95%, and their cell activity was fine and were able to be passaged stably in the subculture. Conclusion It is a successful culture method to extract and purify TSCs, combining primary culture with rapid trypsin digestion with low concentration and differential detachment. After clone screening, the purified and energetic TSCs can be obtained and applied to clinical study for NF2 pathogenesis and treatment. Fibroblasts (FBs) can be removed effectively as well.

DOI：10.3969/j.issn.1672-6731.2011.01.017