The culture and identification of type 1 neurofibromatosis neurofibroma-derived fibroblasts

English Keywords:Type 1 neurofibromatosis  Fibroblast  Cell culture  Tissue culture method

English Abstract:

  【Abstract】 Objective：To explore the method of isolation，culture and identification for type 1 neurofibromatosis（NF-1） neuro fibroma-derived fibroblasts in vitro.Method：The neurofibroma was harvested from four patients with NF-1，and the chipped explants were cultured to obtain the fibroblasts.Differential attachment methoｄ（15 minutes per time）were used to purify the P1 and P2 fibroblasts after the cell overgrew the bottom of the culture flask.The morphology of cultured cells was observed under inverted microscope，immunochemistry was used to test staining of the expression of fibroblast-specific protein-1，vimentin and S-100，ａｎｄ　the fibroblast phenotype　was identified either.Result：Ｌarge number of cultured cells emigrated from the chipped explants 5 days after the primary culture in tissue mass culture method，and 80%-90% confluence of cultured cells was evidenced about 20 days　ｌａｔｅｒ.In the purifｉed P3 fibroblasts，positive staining of fibroblast-specific protein-1 and vimentin and negative staining of S-100 was observed，ｗｉｔｈ　the rate of positive staining　ｏｆ　99.4％ and 99.6％　respectively.Conclusion：The high puriｆｉｅｄ　of NF-1 neurofibroma-derived fibroblasts can be isolated and cultured successfully ｂｙ　tissue mass culture method and differential attachment methoｄ.It ｉｓ an ideal culture system.