深低温冻存软骨细胞构建组织工程化软骨

目的 :研究深低温冻存的软骨细胞作为种子细胞构建组织工程化软骨的能力。方法 :取 2周龄新西兰兔关节软骨细胞 ,经深低温冻存、复苏及体外培养 ,取第 3代对数生长期培养细胞 ,制成细胞悬液 ,调整其为 5× 10 7个 / ml左右 ,接种于 PGA三维支架材料上 ,复合物体外培养 1周 ,移植于裸鼠皮下 ,术后 12周取材 ,行大体及组织学观察。结果 :经深低温冻存的软骨细胞与新鲜的软骨细胞一样形成了预定形态的软骨结构 ,组织学观察有软骨生成及基质分泌 ,单纯PGA支架及单纯的细胞悬液无软骨组织生成。结论 :经深低温冻存的软骨细胞保持了分裂增殖及合成基质的能力 ,可用于组织工程构建组织工程化软骨

Tissue Engineering Cartilage Generated from Cryogenic Chondrocytes

Objective:To study the ability of cryogenically frozen chondrocytes which cultured on a 3 dimensional biodegradable scaffolding material to constructing tissue engieering cartilages.Methods:In this experiment, cryogenically frozen chondrocytes were thawed and cultured in a monolayer in serum based chondrocyte media. They were seeded onto 3 dimensional biopolymer scaffolds in a spinner flask for 1 weeks. The seeded constructs were then transferred to nude mice for 12 weeks. Chondrocyte growth and extracell ular matrix production in the constructs were confirmed by histologic analysis.Results:Histologic sections stained with hematoxylin eosin and Alcian blue (for acidic proteoglycans) and masson(for collagen )confirmed the presence of hyaline cartilage in the cartilage constructs.There was no difference in cryopreserved and noncryopreserved chondrocytes.Conclusions:This study proved that chondrocytes that were cryogenically stored for extended periods could be used to grow cartilage. Cryogenically preserved chondrocytes retained their ability to grow in tissue culture, redifferentiate, and produce extracellular matrix.