| Autor: |
Mithra, Sivaraj, Abdul Majeed, Seepoo, Nafeez Ahmed, Abdul, Suryakodi, Selvam, Rajkumar, Venkatesan, Badhusha, Allahbagash, Kanimozhi, Kumarasamy, Abdul Wazith, Mohamed Jaffer, Taju, Gani, Sahul Hameed, Azeez Sait |
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| Zdroj: |
Aquaculture International; Dec2024, Vol. 32 Issue 7, p9963-9986, 24p |
| Abstrakt: |
Highly susceptible fish cell lines are required to isolate and propagate fish nodavirus and the main purpose of the present study is to develop highly susceptible cell lines from Lates calcarifer which is highly vulnerable to viral nervous necrosis. Three permanent cell lines, brain (SBB), eye (SBE) and optic nerve (SBON) were developed from L. calcarifer by explant culture technique and sub-cultured for 97, 128 and 75 respectively, at 28 °C in Leibovitz's L-15 media supplemented with 10% fetal bovine serum (FBS). The plating efficiency of SBB, SBE and SBON cells was determined as 69.09, 63.19 and 48.22%, respectively, and the doubling time was 27, 29 and 38 h, respectively. All these cell lines were stored by cryopreservation at various passage levels, and a recovery rate of 75–95% was observed. Immunostaining of the SBB and SBON cell lines showed that they are of neuronal origin, whereas the SBE cell line is of retinal pigment epithelial origin. Amplification of mitochondrial 16S rRNA gene confirmed that the SBB, SBE and SBON cell lines were of L. calcarifer origin. Three established cell lines showed 15–18% of transfection efficiency. These cell lines were highly susceptible to striped jack nervous necrosis virus (SJNNV) and infection in these cells was confirmed by RT-PCR, immunocytochemistry, Western blot, ELISA and qRT-PCR. The cell lines established from L. calcarifer were continuous, stable and potentially valuable for virological and biomedical studies. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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