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| Primary cultivation and relative function gene detection of hematocyte in Chinese mitten crab (Eriocheir Sinensis) |
| he 无yang1, 1, 1, 1, 1, |
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Abstract Culture of hemocytes in vitro is of great importance for further molecular study in Chinese mitten crab (Eriocheir Sinensis). In order to investigate optimal culture condition in vitro, such as medium, osmotic pressure and serum concentration, we analyzed cultivation efficiencies of hemocytes in Chinese mitten crab from five different culture medium (1 x L -15, 2 x L-15, 3 x L-15, sf -900 and TC-100 medium), osmotic pressures (original osmotic pressure, 569、803、991、1215 mOsm/kg) and serum concentrations (0%, 5%, 10%, 15%, 20%). In addition, we also detected the relative gene expression in the cultured hemocytes. The result showed that the TC-100 medium was the best according to morphological profiles and survival rate of cultured hemocytes. Furthermore, the best cultivation conditions were determined with osmotic pressure of 991mOsm/kg and 0% bovine serum, which showed the best morphological profile and more than 50% of survival rates after 10 days cultivation. Expression of the three house-keeping gene (β-actin、s-27 and UBE) in Chinese mitten crab were steadily detected in hemocytes in continuously 10 days cultured. In generally, this study indicated that hemocytes cultured in vitro could be used in the further molecular function studies in Chinese mitten crab.
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Received: 16 January 2019
Published: 03 June 2019
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