Hirai Y, Takebe K, Nakajima M, Takashina M, Iizuka M
Biomaterial Research Institute Co., LTD, Yokohama, Japan.
Cytotechnology. 1991 Jul;6(3):209-17. doi: 10.1007/BF00624759.
The functions of hepatocytes under the collagen-contained cell aggregate (cell pack) conditions were studied using liver-specific protein synthesis. Freshly isolated murine hepatocytes were suspended in the medium containing collagen and centrifuged, and the resultant cell masses were cultured on the porous membranes floating on the medium. In these cultures cells were attached to each other three-dimensionally with collagen present in the intercellular spaces. Cultured hepatocytes in the cell pack maintained high and stable activity in the expression of their functions for more than 2 weeks, even when cultured with the medium lacking any hormones and serum, whereas hepatocytes in monolayer cultures lost their functions within a week. Similarly, when the cell packs of rat hepatocytes were transplanted into rat spleens, they could retain viability in the form of cell aggregate with the expression of liver-specific albumin mRNA at a higher level than in the transplanted cell suspensions. The lifespan and the initial expression level of hepatocellular functions in culture were similar to that of the cell pack in cell aggregates without collagen and in cellular monolayers on the collagen gel respectively. It was concluded that the condition where cells are in contact with each other has an important role in the expression of hepatocellular functions and collagen present in the intercellular spaces enhances the functional levels.
利用肝脏特异性蛋白质合成研究了胶原包被的细胞聚集体（细胞包）条件下肝细胞的功能。将新鲜分离的小鼠肝细胞悬浮于含胶原的培养基中并离心，所得细胞团块在漂浮于培养基上的多孔膜上培养。在这些培养物中，细胞通过细胞间存在的胶原三维相互附着。即使在无任何激素和血清的培养基中培养，细胞包中的培养肝细胞在其功能表达方面仍能保持两周以上的高且稳定的活性，而单层培养的肝细胞在一周内就丧失了功能。同样，当将大鼠肝细胞的细胞包移植到大鼠脾脏中时，它们能够以细胞聚集体的形式保持活力，肝脏特异性白蛋白mRNA的表达水平高于移植的细胞悬液。培养中肝细胞功能的寿命和初始表达水平分别与无胶原的细胞聚集体中的细胞包以及胶原凝胶上的细胞单层中的情况相似。得出的结论是，细胞相互接触的条件对肝细胞功能的表达具有重要作用，细胞间存在的胶原可提高功能水平。
