Pollok J M, Ma P X, Vogel C, Dandri M, Petersen J, Burda M R, Kaufmann P M, Kluth D, Rogiers X
Department of Hepatobiliary Surgery, University of Hamburg, Germany.
Dig Surg. 2001;18(3):196-203. doi: 10.1159/000050129.
An optimal method for hepatocyte transplantation is not yet determined. With the principles of tissue engineering in vitro conditioning of hepatocytes on biodegradable polymer in a flow bioreactor before implantation forming spheroids may achieve increased cell mass and function to replace lost organ function in vivo.
Biodegradable poly-L-lactic (PLLA) polymer discs were seeded with rat hepatocytes in a concentration of 10 x 10(6) cells per ml and exposed to a medium flow of 24 ml/min for 1, 2, 4 and 6 days. The number and diameter of spheroidal aggregates was measured by phase-contrast microscopy. H&E histology was performed. Albumin production as hepatocyte specific function was determined by ELISA.
Spheroids of viable hepatocytes of 50-200 microm in diameter were formed. Both the number and diameter of the spheroids increased during the first 2 days and then remained constant until day 6. Albumin production was maintained throughout the culture period.
Short (2- 3 days) pre-transplant conditioning of hepatocytes in a flow bioreactor on biodegradable PLLA resulted in formation of spheroids with a liver-like morphology and preserved specific metabolic function. Tissue engineered hepatocyte spheroids on polymer may represent a functionally active and easy transplantable neotissue and may serve as an in vivo substitute for lost liver function.
肝细胞移植的最佳方法尚未确定。依据组织工程学原理，在植入前于流动生物反应器中，在可生物降解聚合物上对肝细胞进行体外预处理以形成球体，可能会增加细胞数量和功能，从而在体内替代丧失的器官功能。
将大鼠肝细胞以每毫升10×10⁶个细胞的浓度接种于可生物降解的聚-L-乳酸（PLLA）聚合物圆盘上，并使其暴露于流速为24毫升/分钟的培养基中1、2、4和6天。通过相差显微镜测量球状聚集体的数量和直径。进行苏木精-伊红（H&E）组织学检查。通过酶联免疫吸附测定（ELISA）法测定作为肝细胞特异性功能的白蛋白产量。
形成了直径为50 - 200微米的有活力的肝细胞球体。球体的数量和直径在前两天增加，然后直至第6天保持恒定。在整个培养期间白蛋白产量得以维持。
在可生物降解的PLLA上于流动生物反应器中对肝细胞进行短时间（2 - 3天）的移植前预处理，导致形成具有肝样形态且保留特定代谢功能的球体。基于聚合物的组织工程化肝细胞球体可能代表一种功能活跃且易于移植的新组织，并可作为体内丧失肝功能的替代物。
