关键词: 软骨, 培养, 支架, 酶消化法, 生物材料, 组织块培养法, 组织工程

Abstract:

BACKGROUND: Long-segment tracheal resection surgery requires the application of tracheal replacement. The ideal substitute materials should be present with considerable strength, flexibility and inner surface-covered epithelium. Suitable seed cells, excellent carrier, good growth medium are the key components of tissue-engineered trachea construction.
OBJECTIVE: To investigate optimal way for rib chondrocytes isolation and culture, and to construct cartilage-scaffold models.
METHODS: Rabbits’ 5th and 6th rib cartilages were sliced into 1-mm3 pieces and treated with sequential enzyme digest or block culture to isolate chondrocytes. After two passages, rib chondrocytes were collected and suspended. The cell suspension was seeded on poly(lactic acid-glycolic acid) or Dacron scaffold, then cultured under a 37 ℃, 5% CO2 atmosphere. Chondrocytes’ morphology and structure during monolayer culture expansion were observed, while cells growing and matrices formation on scaffold were examined.
RESULTS AND CONCLUSION: Cells isolated by sequential enzyme digest or block culture were primary rib chondrocytes, of which the average passage time was 5 days. However, isolation by tissue block culture was slow and even fibroblasts were found in one flask, indicating that enzyme digest fits for the rib chondrocyte isolation. Chondrocytes attached well on scaffolds. Extracellular matrices were seen to be secreted after 2 weeks. The chondrocytes-scaffold model could be further used to construct tissue-engineered trachea.