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目的 探讨大量分离乳猪肝细胞的方法．方法 用体外灌流装置，ＥＤＴＡ 和胶原酶两步灌流法消化分离乳猪肝细胞，综合方法观察判断肝细胞产量、活力、纯度及增殖能力．结果 分离肝细胞的总产量达到１０９／ 肝，细胞活力９４ ％ ～９８ ％ ，纯度大于９６ ％ ． 分离乳猪肝细胞胞膜完整、细胞器丰富，培养肝细胞有活跃的增殖能力，培养上清中乳酸脱氢酶浓度仅有轻度升高，２４ ｈ 平均浓度１７－８ Ｕ／Ｌ±５－４ Ｕ／Ｌ．结论 采用体外灌流装置两步分离法可获得大量高活性的乳猪肝细胞
Objective To explore a large number of isolated piglets liver cells. Methods The porcine hepatocytes were digested and isolated by two-step perfusion with EDTA and collagenase in vitro, and the hepatocyte production, viability, purity and proliferation were evaluated by comprehensive methods. Results The total yield of isolated hepatocytes reached 109 cells / liver, cell viability 94% -98% and purity> 96%. The isolated suckling liver cells had complete cell membrane, rich organelles, and active proliferation of hepatocytes. The concentration of lactate dehydrogenase in the culture supernatant increased only slightly, and the average concentration of lactate dehydrogenase in the culture supernatant was 17-8 U / L ± 5- 4 U / L. Conclusion A large number of highly active suckling pig hepatocytes can be obtained by the two-step in vitro perfusion device