脂肪来源细胞体外构建组织工程软骨的初步探索

Application of adlpose-derived cells in reconstruction of tissue engineered cartilage in vitro

目的探讨脂肪来源细胞（adipose-derived cells，ADCs）在体外构建特定形态软骨的可行性。方法脂肪组织由整形外科吸脂术获得。酶消化法分离抽吸物中细胞，体外扩增。以第3代细胞接种PLGA生物支架，在成软骨培养基中体外诱导4周，大体观察、组织学检测构建组织的成软骨能力。结果大体观察见诱导组能维持圆柱形态，非诱导组失去原有形态，单纯支架组完全塌陷。组织学上诱导组局部检测到软骨陷窝包埋于嗜碱性基质中，Massons’s染色和Safranin’O染色示胶原、蛋白多糖呈阳性，免疫组织化学染色示Ⅱ型胶原轻度阳性；非诱导组呈典型的疏松结缔组织结构，组织学特殊染色均呈阴性。结论脂肪来源细胞虽为多种细胞混合群体，但作为构建特定形态软骨的种子细胞，具有可行性。

Objective To explore the feasibility of application of adipose-derived cells （ADCs） in reconstruction of tissue engineered cartilage in vitro. Methods Adipose tissue were obtained from human liposuction aspirate （19 cases, 31.5 _＋ 5.8 years old）. ADCs were isolated by collagenase digestion from liposuction aspirates. 3rd passage cells were seeded into PLGA scaffolds. The copolymer constructs were cultured in conditioned or non-conditioned medium in vitro for 4 weeks. The constructs were evaluated though gross morphology, histology, and immunohistochemistry. Results The cell-polymer constructs kept its original shape in the induced group, but lost its original shape in the non-induced group. The scaffold group were collapsed. Histologically, the induced groups showed dense cellularity and lacunae-containing cells embedded in a basophilic matrix, while non-induced groups showed connective tissue-like morphology. Collagen and proteoglycan deposition was revealed by Massons＇s trichome and Safranin＇O staining, and minor collagen H expression in the matrix was detected by immunohistochemistry staining in the induced group. They were all negative in the non- induced groups. Conclusions Although ADCs included many kinds of cells, it is feasible to use ADCs as seeds cells for reconstruction of tissue engineered cartilage.