Isolation and culture of mouse adipose-derived stem cells and their homing to the intestinal tract / 中国组织工程研究

ABSTRACT

BACKGROUND:Adipose-derived mesenchymal stem cels have gained more and more attention due to their high safety, less invasiveness, easy purification and rapid proliferation. OBJECTIVE:To isolate and culture adipose-derived mesenchymal stem cels efficiently and rapidly with high purity and then to explore the cel homing to the intestinal tract after fluorescence labeling. METHODS: Mouse adipose tissue obtained from the groin and epididymis was asepticaly isolated and digested with 0.1% colagenase I. Then the digested cels and undigested adipose tissue were cultured together in the dish to harvest adipose-derived mesenchymal stem cels. The cels were identified by morphology, surface markers, growth kinetics, osteogenic and adipogenic differentiation potential, and then labeled by PKH67 before injected into ulcerative colitis mouse models through the tail vein to observe their homing to the intestinal tract. RESULTS AND CONCLUSION:In vitro, adipose-derived mesenchymal stem cels isolated by this method exhibited spindle-like appearance, grew intensively and arranged in a swirling shape. Adipose-derived mesenchymal stem cels expressed CD29, CD44 and CD90, but not expressed CD45. After osteogenic induction, alkaline phosphatase staining showed black particles and alizarin red S staining showed red mineralized nodules. After adipogenic induction, oil red O staining showed many lipid droplets were dyed red. Cel growth curve showed cels at 3-5 days were in logarithmic growth phase and they were active. Under fluorescence microscopy, frozen sections of the colon were found green fluorescence points that were increased with time. Results suggest that adipose-derived mesenchymal stem cels isolated and culturedin vitro can proliferate rapidly, purify easily and can be induced to osteoblasts and adipocytes; in vivo, PKH67-labeled cels can home to and proliferate in the colon.