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KMID : 0351619740150010097
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Kyungpook Medical Journal
1974 Volume.15 No. 1 p.97 ~ p.104
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Studies on Protein Synthesis of Isolated Liver Cells
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Abstract
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Parenchymal cells abtaired by dispersion of perfused rat 1.ver in 0.25M sucrose solu~:ion were incubated ir. CURL-10EE medium. and their protein. synthesis were studied by measuring the incorporation of amino acid mixed-´~C intJ liver protein.
Adult rat livers were perfused ~~-ith Ca-~ free Locke solution with 0.027i\´I citrate (3;C) and liver cells were separated by passing the mined liver tissue through nylon sieves of increasing fine mesh. The operation was performed b} finer press in 0.25M surcose solution and a sieve pipet cf ´_80 mesh, designed. in cur Iaboratory, ;vas used for final suspension of liver cells. i\~Iajority of the hepatic card cells te.overed iP_ the suspension e~i~red as single ells and the yields were 5~6s 10´ coils from 1 g cf wet Iiver tissue. Liver cells clumps mostly consisted of a fe~.~- but less than ten cells ~~~-ore ai~~~ prepared by incomplete separation and centrifuge method for tl;.e colnparison of the fuac~i~ul with the completly separated liver cell.
Liver cells, suspended in C:~IRLlOG6 mediuri 15 percent rat serum, were transfe-ed to th;; incubation. apparatus (Fig i) and the cells were reaggregated in the medium or figred on the inner surface of incubation tubes by centroIing the speed of tube rotation within 30 mi_~utes after initiation cf coil inc~b~tiors.
The liver cells of two different condition during 4 hours incuba~ien showed a considerable difference in the incorporation stud}r. In filed liver cells more increase of the incorporation was noted after 2 hours whereas the incorporation was aImest stopped is reaggregated cells after the time.
Considering the possible effect of exposure of liver cells in to air phase in the filed state acc^rding to t?-~e tube rotation, etperiement replaced the air phase partialy with oxygen was also carried gut, but no :.oticeable effect of additional oxygen supply on the incorporation was noted.
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