Takabatake H, Koide N, Tsuji T
First Department of Internal Medicine, Okayama University Medical School, Japan.
Artif Organs. 1991 Dec;15(6):474-80.
Multicellular spheroids are spherical cell-aggregates that retain tridimensional architecture and tissue-specific functions. For use of multicellular spheroids of hepatocytes in a bioreactor for hybrid artificial liver support, we studied the effect of encapsulation and circulating culture on their integrity and tissue-specific functions. Multicellular spheroids of rat hepatocytes were encapsulated into microdroplets of calcium alginate gel and were used as a bioreactor in medium circulating in a spouted bed chamber. Approximately 10% of the hepatocytes of an adult rat were entrapped in a bioreactor chamber, connected to a gas exchanger and a medium reservoir. The total bed volume of the system was 250 ml. The pH and DO2 of the hormonally defined circulating medium was maintained constantly. Albumin and urea were produced in a linear fashion for 64 h at the rates of 0.02 micrograms/microgram cell protein/day and 0.15-0.2 ng/micrograms cell protein/day, respectively. Viability and structural stability of the spheroids were well preserved after the culture period. These results indicate that these encapsulated multicellular hepatocyte spheroids will provide a useful bioreactor for the continuous production of albumin, in vitro and also a prototype hybrid artificial liver support.
多细胞球体是保留三维结构和组织特异性功能的球形细胞聚集体。为了在用于混合人工肝支持的生物反应器中使用肝细胞多细胞球体，我们研究了包封和循环培养对其完整性和组织特异性功能的影响。将大鼠肝细胞多细胞球体包封到海藻酸钙凝胶微滴中，并用作在喷动床腔室中循环的培养基中的生物反应器。将约10%的成年大鼠肝细胞捕获在与气体交换器和培养基储存器相连的生物反应器腔室中。该系统的总床体积为250毫升。激素定义的循环培养基的pH值和溶解氧水平保持恒定。白蛋白和尿素分别以0.02微克/微克细胞蛋白/天和0.15 - 0.2纳克/微克细胞蛋白/天的速率呈线性产生64小时。培养期后球体的活力和结构稳定性得到良好保存。这些结果表明，这些包封的多细胞肝细胞球体将为体外连续生产白蛋白提供有用的生物反应器，也是混合人工肝支持的原型。
