复合胶原纳米粒子在猪肝细胞培养中的应用

目的:制备聚乳酸-O-羧甲基壳聚糖(PLA-O-CMC)纳米材料和Ⅰ型胶原复合物进行猪肝细胞的体外培养。方法:用超声乳化法制备PLA-O-CMC纳米粒子,扫描电镜对其进行表征。采用Ⅳ型胶原酶原位循环灌流法分离猪肝细胞。将肝细胞分为:Ⅰ组(裸细胞培养组)、Ⅱ组(复合胶原纳米粒子细胞混合培养组)进行培养,观察肝细胞形态结构及功能变化。结果:PLA-0-CMC纳米粒子及Ⅰ型胶原为肝细胞的生长提供了良好的支架和外环境,体外肝细胞的生长也从平面型向三维型迈出了较大的一步。培养猪肝细胞能保持良好的活性和白蛋白分泌功能。结论:PLA-0-CMC纳米粒子及Ⅰ型胶原体为肝细胞生长的良好载体。

The preparation of nanoparticles and collagen Ⅰcompound in culture of porcine hepatocytes

Objective: To prepare polymer of lactic acid-O-carboxymethylated chitosan(PLA-O-CMC) nanomaterials and collagen Ⅰ for the cultivation of porcine hepatocytes in vitro. Methods: Polylatic acid-O-carboxymethylated chitosan (PLA-O-CMC) nanoparticles were prepared with PLA and O-CMC by ultrasonic method. Nanoparticles were characterized with x-ray photoelectron spectrometry. Porcine hepatocytes were isolated by in situ recirculating collagenaseIV perfusion mothed. Morphological, vitality, albumin and activity of hepatocytes growing in Group Ⅰ(common culture group), Group Ⅱ(co-cultured hepatocytes group with collagen Ⅰand PLA-O-CMC nanoparticles) were observed, respectively. Results: The compounds of PLA-O-CMC nanomaterials and collagen Ⅰ profit the aggregation of hepatocytes cultured in vitro and enhance their biological function, which could satisfy the need for the number of hepatocytes by construction of the liver tissue engineering essentially.