Chief EditorK.M.L. Pathak
Print ISSN 0367-6722
Online ISSN 0976-0555
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Five Layered Optiprep based Density Gradient Model is a Promising Model for Enrichment of Viable X Chromosome Bearing Spermatozoa in Bubalus bubalis
First Online 22-08-2020|
Methods: Density gradient centrifugation (DGC) technique was employed for enriching X chromosome spermatozoa in buffalo bull semen using four gradient media viz., Percoll, Optiprep, Ficoll and Sucrose. Percentage enrichment of X chromosome bearing spermatozoa in all the DGC models was determined by SYBR green based Real Time PCR.
Result: Our investigations revealed that number of layers and centrifugation speed (g) factors in a density gradient centrifugation model, have significant effect on the percentage enrichment of X chromosome content in semen samples.Three layers, four layers and five layers density gradient centrifugation models showed significant differences (P<0.05) in the sex ratio towards X spermatozoa population in semen (61.72%±0.81, 64.55%±0.26 and 67.31±0.33%) respectively. Density gradient centrifugation models with centrifugation speed (g) 200x g and 300x g also showed significant differences (P<0.05) in the sex ratio towards X spermatozoa population in semen (66.36%±0.25 and 62.69±0.22%) respectively. Out of all DGC models, Optiprep density gradient model with 5 layers and centrifugation at 200xg (O3) yielded maximum enrichment (72.4%±1.38). In conclusion, O3 poses to be a promising model for enrichment of X spermatozoa in buffalo bull semen.
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