Comparative Study on Enzymatic and Explant Method in Establishing Primary Culture from Different Cultivable Cells of Indian Major Carp, Cirrhinus mrigala

摘要

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.