Hepatic differentiation of canine and human induced pluripotent stem cells (iPS cells)
Grauw, N. de
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The aim of this study was to look at the possibilities whether canine and human induced pluripotent stem cells (iPS cells) can be differentiated into hepatocytes. And if this differentiation was possible we also looked if the cultured hepatocytes were functionally similar to primary hepatocytes. During this study we created an optimized, cost-effective, protocol for hepatic differentiation of iPS cells based on previous studies. This protocol imitates the embryonic development. To demonstrate the differentiation towards hepatocytes with all intermittent steps, gene-expression, immunocytochemistry and functional tests were performed; qPCR, immunocytochemistry, indocyanine green staining, Periodic Acid Schiff staining, liver transaminase determination, Cytochrome P450 test, micro albumin determination and ammonia tolerance test. The results of both of the cell lines indicate that the cells are similar, only the human hepatocytes seem to have a higher expression of hepatocyte specific genes. The cultured iPS cell derived hepatocytes seem to look similar in function with primary hepatocytes according to all the tests we performed. Not all test results are entirely convincing and some test should be performed once again to achieve better test results (cytochrome test, ammonia tolerance test).