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Articles by S.K. Nayak
Total Records ( 2 ) for S.K. Nayak
  P.K. Nanda , P. Swain , S.K. Nayak , T. Behera and K. Dhama
  Enzymatic method using trypsin to establish primary culture from various cells from heart, liver, kidney, brain, testis, ovary, fin and embryo of Indian Major Carp, Cirrhinus mrigala was investigated and compared with that of explant culture method. The dissociation time to obtain cells from individual tissues and the viability of cells for primary culture varied at 0.125 and 0.25% trypsin concentration. Single cell culture of trypsinised cells from testis, ovary and embryo showed good attachment irrespective of trypsin concentration. Cells from liver and heart were found to be sensitive to trypsin only at higher concentration (0.25%); whereas, no attachment/or proliferation was observed from cells of kidney, brain and fin due to sensitivity to trypsin at both the concentrations. In contrast to this method to obtain primary culture, explant culture from most of the tissues showed better attachment, resulting in subsequent growth and proliferation of cells forming monolayer. Overall, the explant culture of most of the tissues of C. mrigala was found to be suitable and survived more passages as compared to single cell culture obtained through trypsinisation. For obtaining primary culture from single cells, further investigations are desired to identify tissue specific enzymes, standardization of dose, duration and temperature of enzymatic treatment.
  P.K. Nanda , P. Swain , S.K. Nayak , T. Behera , P. Jayasankar and K. Dhama
  The success in establishing in vitro culture from anchorage dependent tissue explants depends upon their ability to attach to culture substratum. In this study, evaluation of different coating factors on the attachment and subsequent monolayer formation from various tissue explants of Indian Major Carp, Mrigal, Cirrhinus mrigala was investigated to establish cell culture. Explants from heart, liver, gills, testis, ovary, kidney and fin were taken in respective culture flasks coated either with type I collagen, gelatin or fibronectin to know their ability on attachment and proliferation of cells resulting in attaining confluency. The percentage attachment of explants from same tissue, when cultured in different coated flasks, varied greatly. Explants (40-65%) from liver, ovary, testis and fin were found to attach well in fibronectin coated flasks, whereas maximum attachment (>65%) from heart tissue was recorded in gelatin coated flasks. This difference in percentage attachment of explants to substratum significantly affected growth, proliferation of cells and subsequent formation of confluent monolayer. Overall, fibronectin as a coating factor was found to be good for attachment of explants from most of the tissues of C. mrigala. Contrary to this, attachment percentage of explants from fin, gills, ovary and testis in type I collagen coated flasks was less which eventually failed to form monolayer. The findings indicate that coating factor(s) has a major role in establishing cell culture which not only influences attachment but also growth and proliferation of cells from explants obtained from different tissues of C. mrigala. Further investigations are suggested to find out suitable coating factors for each cell types of different fish species so as to establish cell cultures.
 
 
 
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