关键词: 绿色荧光蛋白, 转基因小鼠, 骨髓间充质干细胞, 多向分化潜能

Abstract:

Objective　To study the multi-lineage potential of bone marrow mesenchymal stem cells (MSCs) derived from transgenic mice with green fluorescent protein (GFP) genein vitro.Methods　A6-week-old GFP transgenic mouse was executed by dislocation of cervical vertebra, and the marrow in tibia and thighbone was washed outwith asepsis. The limited cell strains of MSCs derived from GFP transgenic mice(GFP-MSCs)were obtained with density gradient centrifugation. The passage 3 GFP-MSCs were induced to differentiate into osteoblast, adippcyte, neuron with solution of calcium induction medium, adipogenic medium and neural induction medium respectively. After being calcium-induced, the activity of alkaline phosphatase on GFP-MSCs was determined by micro-plate reader, and alizarin red staining was performed to test the formation of calcium concentration. The adipo-induced MSCs were detected with oil red O staining. Immunocytochemical stainingwas performed to detect the expression of NSE on neuron-induced MSCs.Results　The ALP activity of GFP-MSCs heightened gradually alongwith being calcium-induced, and alizarin red staining showed positive. Oil red O staining of adipo-induced cells and NSE immunocytochemical staining of neu- ron-induced cells demonstrated positive.Conclusion　The limited cell strain of GFP-MSCs possessesmulti-lineage potential,which can be used as an efficient tracking facility for studying the mechanism of multi-lineage potential on the MSCs.

Key words: green fluorescent protein, transgenic mice, mensenchymal stem cells, multi-lineage potential