Xiong M, Ai Y F, Wang Y
Plastic Surgery Center, XiJing Hospital, Fourth Military Medical University, Xi'an Shanxi, P. R. China 710032.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2001 Mar;15(2):109-12.
To explore the feasibility of reconstructing tissue engineered vessel in vitro.
Bovine endothelial cells were isolated from calf thoracic aorta by enzyme digestion methods and subcultured and purified. The endothelial cells of the 3rd to 7th passages were seeded into the inner surface of tubular scaffold material by polyglycolic acid(PGA) coated with cross-linked collagen, and cultured in vitro for 10 days using dynamic rotation culture technique. Scanning electron microscopy was used to analyse the morphological characteristics, and prostacyclin released by endothelial cells was measured by radioimmunoassay of 6-keto-prostaglandin F1 alpha.
The VIII factor staining of cultured endothelial cells was positive. The endothelial cells adhered well on the inner surface of tubular scaffold material with confluent monolayer covering(91.2 +/- 1.5)%. The endothelialized model released prostacyclin at a rate of (4.6 +/- 0.5) micrograms/cm2.min. There was significant difference to control group (P < 0.05).
The PGA coating with collagen is an ideal scaffold for endothelial cells, the coverage rate is increased through dynamic rotation culture technique. It will lay a good foundation for architecture of a laminated structure of tissue engineered vessel.
探讨体外构建组织工程血管的可行性。
采用酶消化法从小牛胸主动脉分离牛内皮细胞,进行传代培养及纯化。将第3至7代内皮细胞接种于交联胶原包被的聚乙醇酸(PGA)管状支架材料内表面,采用动态旋转培养技术体外培养10天。用扫描电子显微镜分析其形态学特征,通过放射免疫法测定6-酮-前列腺素F1α来检测内皮细胞释放的前列环素。
培养的内皮细胞Ⅷ因子染色呈阳性。内皮细胞在管状支架材料内表面黏附良好,汇合单层覆盖率为(91.2±1.5)%。内皮化模型释放前列环素的速率为(4.6±0.5)μg/cm²·min,与对照组相比有显著差异(P<0.05)。
胶原包被的PGA是内皮细胞理想的支架材料,通过动态旋转培养技术可提高覆盖率,为构建组织工程血管的分层结构奠定良好基础。