Rapid, large-scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver
Rapid, large-scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver
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DOI：
10.1002/lt.20446
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发表时间：
2005-08-01
影响因子：
4.6
通讯作者：
Rinaldo, P
中科院分区：
文献类型：
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作者：
Nyberg, SL;Hardin, J;Rinaldo, P
我们已经开发了一种新型的生物反应器的基础上观察，分离的猪肝细胞迅速自发聚集成球状体振荡条件下。本研究的目的是表征振荡频率（0.125 Hz，0.25 Hz）、细胞密度（1-10 X 10（6）个细胞/mL）和储存条件（新鲜、冻存）对猪肝细胞球状体形成动力学的影响。还将球状体肝细胞的活力和代谢性能与单层培养进行了比较。我们观察到新鲜和冻存的猪肝细胞在振荡培养开始时都自发地开始形成球状体。球体大小与细胞密度和培养时间直接相关，但与振荡频率呈负相关。新鲜猪肝细胞的球状体形成与细胞死亡减少有关（乳酸脱氢酶释放，1.3 +/- 1.0 vs. 3.1 +/- 0.7 U/mL，P < 0.05）和代谢性能增加（白蛋白产生，14.7 +/- 3.3 vs.4.6 +/- 1.4 fg/c/h，P < 0.0001;氨的尿素生成，267 +/- 63 vs.92 +/- 13 lAmol/ L/h，P < 0.001）。总之，基于快速球状体形成、肝细胞功能增加和扩大规模的有利特性，球状体储库生物反应器值得进一步研究作为支持肝功能衰竭的生物人工肝。
We have developed a novel bioreactor based on the observation that isolated porcine hepatocytes rapidly and spontaneously aggregate into spheroids under oscillation conditions. The purpose of this study was to characterize the influence of oscillation frequency (0.125 Hz, 0.25 Hz), cell density (1-10 X 10(6) cells/mL), and storage condition (fresh, cryopreserved) of porcine hepatocytes on the kinetics of spheroid formation. The viability and metabolic performance of spheroid hepatocytes was also compared to monolayer culture. We observed that both fresh and cryopreserved porcine hepatocytes began formation of spheroids spontaneously at the onset of oscillation culture. Spheroid size was directly related to cell density and time in culture, though inversely related to oscillatory frequency. Spheroid formation by fresh porcine hepatocytes was associated with decreased cell death (lactate dehydrogenase release, 1.3 +/- 1.0 vs. 3.1 +/- 0.7 U/mL, P < 0.05) and increased metabolic performance (albumin production, 14.7 +/- 3.3 vs. 4.6 +/- 1.4 fg/c/h, P < 0.0001; ureagenesis from ammonia, 267 +/- 63 vs. 92 +/- 13 lAmol/ L/h, P < 0.001) compared with monolayer culture. In conclusion, based on the favorable properties of rapid spheroid formation, increased hepatocellular function, and case of scale-up, the spheroid reservoir bioreactor warrants further investigation as a bioartificial liver for support of liver failure.