Kinasiewicz Andrzej, Smietanka Anna, Dudziński Konrad, Chwojnowski Andrzej, Gajkowska Barbara, Weryński Andrzej
Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland.
Artif Organs. 2008 Sep;32(9):747-52. doi: 10.1111/j.1525-1594.2008.00600.x. Epub 2008 Jul 30.
There are different types of membranes used for hepatocyte cultivation. In our studies, spongy polyethersulfone (PES) membranes were examined as a support for hepatic cell cultivation in vitro. The extended surface of the membranes allows to introduce a high cell number especially in three-dimensional gel structure. Scanning electron microscopy analysis indicated that C3A cells used in our experiments grew well on PES membranes forming microvilli characteristic for normal hepatocytes. Analysis of cell viability proved that spongy PES membrane is well tolerated by J774 macrophages and did not stimulate nitric oxide synthesis. Bile canalicular structures were observed in fluorescence microscopy after F-actin staining with tetramethyl rhodamine iso-thiocyanate (TRITC)-phalloidin. The C3A cells showed high affinity to the PES membranes and adhered to almost 90% during the initial 24 h of incubation. Albumin production increased during static culture from the value of 805.2 +/- 284.4 (ng/24 h/initial 10(6) cells) during the first days, to 2017.6 +/- 505.9 (ng/24 h/initial 10(6) cells) after 10 days of culture. In conclusion, the spongy PES membranes can be used as scaffold for hepatocyte cultivation, especially for the creation of three-dimensional environments.
用于肝细胞培养的膜有不同类型。在我们的研究中，对海绵状聚醚砜（PES）膜进行了检测，以作为体外肝细胞培养的支撑物。这些膜的扩展表面能够引入大量细胞，特别是在三维凝胶结构中。扫描电子显微镜分析表明，我们实验中使用的C3A细胞在PES膜上生长良好，形成了正常肝细胞特有的微绒毛。细胞活力分析证明，海绵状PES膜能被J774巨噬细胞良好耐受，且不会刺激一氧化氮的合成。在用异硫氰酸四甲基罗丹明（TRITC）-鬼笔环肽进行F-肌动蛋白染色后，在荧光显微镜下观察到胆小管结构。C3A细胞对PES膜表现出高亲和力，在孵育的最初24小时内，附着率几乎达到90%。在静态培养过程中，白蛋白产量从最初几天的805.2±284.4（ng/24小时/初始10⁶个细胞）增加到培养10天后的2017.6±505.9（ng/24小时/初始10⁶个细胞）。总之，海绵状PES膜可作为肝细胞培养的支架，特别是用于创建三维环境。
