Study of the effect of microcapsule fluidized culture on C3A cell function and its molecular mechanism
LI Xiaoqing1 LI Jianzhou2,3 YANG Ying3 LI Lanjuan3
1.Department of Emergency, First Affiliated Hospital of Xi’an JiaoTong University, Shaanxi Province, Xi’an 710061, China;
2.Department of Infectious Diseases, First Affiliated Hospital of Xi’an JiaoTong University, Shaanxi Province, Xi’an 710061, China;
3.State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Zhejiang Province, Hangzhou 310003, China
Abstract Objective To investigate the effect of microcapsule fluidized culture on C3A cell function and its molecular mechanism. Methods Base on the different culture modes of C3A cells, they were divided into three groups: two-dimensional culture group, microcapsule static culture group and microcapsule fluidized culture group. The effect of microencapsulation fluidized culture on the activity of C3A cells, albumin, urea synthesis capacity and metabolism ability was evaluated. The expression level of genes related to hepatocyte function was detected. The expression levels of key molecules in metabolism-related signaling pathways of hepatocytes were detected. Results The albumin, urea synthesis capacity and CYP1A2, CYP3A4 metabolic activity of C3A cells in the microcapsule static culture group and microencapsulation fluidized culture group were higher than those in the two-dimensional culture group, and the differences were statistically significant (P ＜ 0.05). Compared with the microcapsule static culture group, the albumin, urea synthesis capacity and CYP1A2 metabolic activity of C3A cells in the microcapsule fluidized culture group were increased, and the differences were statistically significant (P ＜ 0.05). The expression levels of CYP1A2, CYP2B62, CYP2E1, CYP3A4 and UGT2B4 genes of C3A cells in the microcapsule static culture group and microencapsulation fluidized culture group were higher than those in the two-dimensional culture group, and the differences were statistically significant (P ＜ 0.05). Compared with the microcapsule static culture group, the expression levels of p-PKC, p-PKA and calmodulin (CaM) proteins related to metabolism of C3A cells in the microcapsule fluidized culture group were decreased, and the differences were statistically significant (P ＜ 0.05). Conclusion Microcapsule fluidized culture can significantly improve the synthesis and metabolism fuction of C3A cells, and its mechanism of action may be related to the down-regulation of the phosphorylation level of related signaling pathway proteins and level of CaM.
LI Xiaoqing1 LI Jianzhou2,3 YANG Ying3 LI Lanjuan3. Study of the effect of microcapsule fluidized culture on C3A cell function and its molecular mechanism[J]. 中国医药导报, 2021, 18(15): 4-8,26.
[1] Sussman NL，Kelly JH. Artificial liver [J]. Clin Gastroenterol Hepatol，2014，12（9）：1439-1442.
[2] Larsen FS. Artificial liver support in acute and acute-on-chronic liver failure [J]. Curr Opin Crit Care，2019，25（2）：187-191.
[3] Patel P，Okoronkwo N，Pyrsopoulos NT. Future Approaches and Therapeutic Modalities for Acute Liver Failure [J]. Clin Liver Dis，2018，22（2）：419-427.
[4] Duan Z，Xin S，Zhang J，et al. Comparison of extracorporeal cellular therapy ELAD（R）vs standard of care in a randomized controlled clinical trial in treating Chinese subjects with acute-on-chronic liver failure [J]. Hepat Med，2018，10：139-152.
[5] Thompson J，Jones N，Al-Khafaji A，et al. Extracorporeal cellular therapy（ELAD）in severe alcoholic hepatitis：A multinational，prospective，controlled，randomized trial [J]. Liver Transpl，2018，24（3）：380-393.
[6] Ocskay K，Kanjo A，Gede N，et al. Uncertainty in the impact of liver support systems in acute-on-chronic liver failure：a systematic review and network meta-analysis [J]. Ann Intensive Care，2021，11（1）：10.
[7] Ng SS，Xiong A，Nguyen K，et al. Long-term culture of human liver tissue with advanced hepatic functions [J]. Jci Insight，2017，2（11）：e90853.
[8] Hu H，Gehart H，Artegiani B，et al. Long-Term Expansion of Functional Mouse and Human Hepatocytes as 3D Organoids [J]. Cell，2018，175（6）：1591-1606.
[9] Messner S，Agarkova I，Moritz W，et al. Multi-cell type human liver microtissues for hepatotoxicity testing [J]. Arch Toxicol，2013，87（1）：209-213.
[10] Serp D，Cantana E，Heinzen C，et al. Characterization of an encapsulation device for the production of monodisperse alginate beads for cell immobilization [J]. Biotechnol Bioeng，2000，70（1）：41-53.
[11] He YT，Qi YN，Zhang BQ，et al. Bioartificial liver support systems for acute liver failure：A systematic review and meta-analysis of the clinical and preclinical literature [J]. World J Gastroenterol，2019，25（27）：3634-3648.
[12] Zhao LF，Pan XP，Li LJ. Key challenges to the development of extracorporeal bioartificial liver support systems [J]. Hepatobiliary Pancreat Dis Int，2012，11（3）：243-249.
[13] Pan XP，Li LJ. Advances in cell sources of hepatocytes for bioartificial liver [J]. Hepatobiliary Pancreat Dis Int，2012，11（6）：594-605.
[14] Gu C，Chai M，Liu J，et al. Expansion of Transdifferentiated Human Hepatocytes in a Serum-Free Microcarrier Culture System [J]. Dig Dis Sci，2020，65（7）：2009-2023.
[15] Yan S，Wei J，Liu Y，et al. Hepatocyte spheroid culture on fibrous scaffolds with grafted functional ligands as an in vitro model for predicting drug metabolism and hepatotoxicity [J]. Acta Biomater，2015，28：138-148.
[16] Ito H，Kamiya A，Ito K，et al. In vitro expansion and functional recovery of mature hepatocytes from mouse adult liver [J]. Liver Int，2012，32（4）：592-601.
[17] Sarika PR，James NR，Anilkumar PR，et al. Microgravity as a means to incorporate HepG2 aggregates in polysaccharide-protein hybrid scaffold [J]. J Mater Sci Mater Med，2016，27（2）：27.
[18] Ruoss M，Vosough M，Konigsrainer A，et al. Towards improved hepatocyte cultures：Progress and limitations [J]. Food Chem Toxicol，2020，138：111188.
[19] He Y，Liu C，Xia X，et al. Conformal microcapsules encapsulating microcarrier-L02 cell complexes for treatment of acetaminophen-induced liver injury in rats [J]. J Mater Chem B，2017，5（10）：1962-1970.
[20] Ozawa F，Ino K，Arai T，et al. Alginate gel microwell arrays using electrodeposition for three-dimensional cell culture [J]. Lab Chip，2013，13（15）：3128-3135.
[21] Chen Y，Yu C，Lv G，et al. Rapid large-scale culturing of microencapsulated hepatocytes：a promising approach for cell-based hepatic support [J]. Transplant Proc，2014，46（5）：1649-1657.
[22] Chan HF，Zhang Y，Leong KW. Efficient One-Step Production of Microencapsulated Hepatocyte Spheroids with Enhanced Functions [J]. Small，2016，12（20）：2720-2730.
[23] Dhawan A，Chaijitraruch N，Fitzpatrick E，et al. Alginate microencapsulated human hepatocytes for the treatment of acute liver failure in children [J]. J Hepatol，2020，72（5）：877-884.
[24] Boon R，Kumar M，Tricot T，et al. Amino acid levels determine metabolism and CYP450 function of hepatocytes and hepatoma cell lines [J]. Nat Commun，2020，11（1）：1393.
[25] Zanger UM，Schwab M. Cytochrome P450 enzymes in drug metabolism：regulation of gene expression，enzyme activities，and impact of genetic variation [J]. Pharmacol Ther，2013，138（1）：103-141.
