Establishment of gene-edited pigs expressing human blood-coagulation factor VII and albumin for bioartificial liver use
Li Li
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorHongyi Meng
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorQingjian Zou
School of Chemical and Environmental Engineering, Wuyi University, Jiangmen, China
Search for more papers by this authorJianmin Zhang
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorLei Cai
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorBin Yang
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorJun Weng
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Liangxue Lai
South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Huaqiang Yang
College of Animal Science, South China Agricultural University, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Yi Gao
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorLi Li
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorHongyi Meng
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorQingjian Zou
School of Chemical and Environmental Engineering, Wuyi University, Jiangmen, China
Search for more papers by this authorJianmin Zhang
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorLei Cai
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorBin Yang
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorJun Weng
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Liangxue Lai
South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Huaqiang Yang
College of Animal Science, South China Agricultural University, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Yi Gao
Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Correspondence
Dr Huaqiang Yang, College of Animal Science, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
Email: [email protected]
Dr Yi Gao, Department of Hepatobiliary Surgery II, Zhujiang Hospital, Southern Medical University, 253 Industry Avenue, Guangzhou 510280, China.
Email: [email protected]
Dr Liangxue Lai, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Avenue, Guangzhou 510530, China.
Email: [email protected]
Search for more papers by this authorAbstract
Background and Aim
Bioartificial livers (BALs) are considered as a solution to bridge patients with acute liver failure to liver transplantation or to assist in spontaneous recovery for patients with end-stage liver disease. Pig is the best donor of hepatocytes for BALs in clinical trials, because metabolic and detoxification function of its liver are close to human. However, using pig hepatocytes for BALs remains controversial for safety concern owing to nonhuman proteins secretion. Herein, we attempt to establish modified pigs expressing humanized liver proteins, blood-coagulation factor VII (F7), and albumin (ALB). These pigs should also be porcine endogenous retrovirus subtype C (PERV-C) free so that their ability of transmitting PERV to human could be diminished seriously.
Methods
We devised both homology-dependent and independent knock-in approaches to insert a fusion of hF7 and hALB gene downstream the site of pig endogenous F7 promoter in pig fetal fibroblasts negative for PERV-C. The modified pigs were then generated through somatic cell nuclear transfer.
Results
We obtained 14 and 10 cloned pigs by homology-dependent and independent approaches, respectively. Among them, 19 cloned pigs were with expected gene modification and 13 are alive to date. These modified pigs can successfully express hF7 and hALB in the liver and serum, and the expressed hF7 exhibits normal coagulation activity.
Conclusions
The gene-edited pigs expressing hF7 and hALB in the liver were generated successfully. We anticipate that our pigs could provide an alternative cell source for BALs as a promising treatment for patients with acute liver failure.
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