P-242

Benefit of Islets on Hepatocyte in Co-Culture

C.-H. Kuo¹, H.-L. Peng¹, J.-H. Juang^2
1 National Chiao Tung Unversity, The Department of Biological Science and Technology, Hsinchu , Taiwan; ² Chang Gung University and Chang Gung Memorial Hospital, Taoyuan, Taiwan

Although the Liver transplantation techniques are well established, the irreversibility of the procedure, high cost, and scarcity of donor organs are some of the remaining problem. The advantage of hepatocyte transplantation is the possibility of cell culture, which allows independence from the limitations by ischemia of donated organs.

Objective: We investigated the effect of islets on hepatocyte.

Methods: Hepatocyte and islets were freshly isolated from the male C57BL/6J mice. Hepatocyte were cultured in collagen type I gel as a group A (control), and the separated co-culture with islets as a group B. In group A: 10⁵ hepatocyte , Group B: 10⁵ hepatocyte separated co-culture with 150 islets. We examined cell variability with MTT, and checked cell apoptosis with Annexin V and PI (double staining) in flow cytometry. We also examed the albumin secretion by enzyme-linked immunoadsorbent assay on day 1,3,5,7.

Results: We found the hepatocyte cannot resist the isolated condition from organ or tissue. Hepatocyte hurriedly necrosis and albumin secretion reduced. But, on Day 7 in group B HSCs still maintain the well function. Albumin secretion Group A:0.1451 ± 0.0329 pg/ml/cell vs. 0.3351 ± 0.0581 pg/ml/cell, p value<0.05. The necrosis and apoptosis sigmal data are, apoptosis sections: day 1, Group A: 9.90% group B:7.52%; day 3, Group A:13.85%, group B:7.62%. Necrosis: day 1, Group A: 27.92% group B:15.29%; day 3 30.31%, group B:20.40%..

Conclusions: These findings, maybe islets provide the cell survival factor for hepatocyte in vitro. Our studies, both cells separate from the membrane, the effects of the islets were mediated by soluble factors. We observes the result islets are benefit on hepatocyte sotrage before hepatocyte transplantation in the future. This co-culture system, which incorporates hepatocytes and islets, is considered to be important in the development of high-performance bioartificial liver (BAL) system.

Parallel poster session 6 - Topic: Immunoisolation and bioartificial pancreas

P-242

Benefit of Islets on Hepatocyte in Co-Culture

C.-H. Kuo¹, H.-L. Peng¹, J.-H. Juang^2
1 National Chiao Tung Unversity, The Department of Biological Science and Technology, Hsinchu , Taiwan; ² Chang Gung University and Chang Gung Memorial Hospital, Taoyuan, Taiwan

Although the Liver transplantation techniques are well established, the irreversibility of the procedure, high cost, and scarcity of donor organs are some of the remaining problem. The advantage of hepatocyte transplantation is the possibility of cell culture, which allows independence from the limitations by ischemia of donated organs.

Objective: We investigated the effect of islets on hepatocyte.

Methods: Hepatocyte and islets were freshly isolated from the male C57BL/6J mice. Hepatocyte were cultured in collagen type I gel as a group A (control), and the separated co-culture with islets as a group B. In group A: 10⁵ hepatocyte , Group B: 10⁵ hepatocyte separated co-culture with 150 islets. We examined cell variability with MTT, and checked cell apoptosis with Annexin V and PI (double staining) in flow cytometry. We also examed the albumin secretion by enzyme-linked immunoadsorbent assay on day 1,3,5,7.

Results: We found the hepatocyte cannot resist the isolated condition from organ or tissue. Hepatocyte hurriedly necrosis and albumin secretion reduced. But, on Day 7 in group B HSCs still maintain the well function. Albumin secretion Group A:0.1451 ± 0.0329 pg/ml/cell vs. 0.3351 ± 0.0581 pg/ml/cell, p value<0.05. The necrosis and apoptosis sigmal data are, apoptosis sections: day 1, Group A: 9.90% group B:7.52%; day 3, Group A:13.85%, group B:7.62%. Necrosis: day 1, Group A: 27.92% group B:15.29%; day 3 30.31%, group B:20.40%..

Conclusions: These findings, maybe islets provide the cell survival factor for hepatocyte in vitro. Our studies, both cells separate from the membrane, the effects of the islets were mediated by soluble factors. We observes the result islets are benefit on hepatocyte sotrage before hepatocyte transplantation in the future. This co-culture system, which incorporates hepatocytes and islets, is considered to be important in the development of high-performance bioartificial liver (BAL) system.

Parallel poster session 6 - Topic: Immunoisolation and bioartificial pancreas