Replacement of serum with ocular fluid for cryopreservation of immature testes.
Cryopreservation of immature testis is a feasible approach for germplasm preservation of male animals. Combinations of dimethyl sulfoxide (DMSO) and foetal bovine serum (FBS) are used for testis cryopreservation. However, an alternative to FBS is needed, because FBS is expensive. Buffalo ocular fluid (BuOF), a slaughter house by-product, could be an economical option. The objective of the present study was to assess whether BuOF can replace FBS for cryopreservation of immature mouse (Mus musculus), rat (Rattus norvegicus), and buffalo (Bubalus bubalis) testes. Results showed that rodent and buffalo testes frozen in DMSO (10% for rodents and 20% for buffalo) with 20% FBS or BuOF had similar numbers of viable and DNA-damaged cells (P > 0.05). The expression of cell proliferation- (PCNA) and apoptosis-specific proteins (Annexin V and BAX/BCL2 ratio) were also comparable in mouse and buffalo testes frozen in DMSO with FBS or BuOF (P > 0.05). Interestingly, rat testis frozen in DMSO with BuOF had lower expression of Annexin V protein than testis frozen in DMSO with FBS (P < 0.05). The percentage of meiotic germ cells (pachytene-stage spermatocytes) in xenografts from testis frozen either in DMSO with BuOF or FBS did not significantly differ in rats or buffalo (P > 0.05). These findings provide evidence that BuOF has potential to replace FBS for cryopreservation of immature rodent and buffalo testis. Further investigation is needed to explore whether BuOF can replace FBS for testis cryopreservation of other species.
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Lavanya
L
Pothana
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Lalitha
L
Devi
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Naresh Kumar
NK
Venna
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Niharika
N
Pentakota
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Vivek Phani
VP
Varma
Animal House, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
Jedy
J
Jose
Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India. Electronic address: sandeep@ccmb.res.in.
Sandeep
S
Goel
D000818:Animals; D001826:Body Fluids; D002020:Buffaloes; D002417:Cattle; D049109:Cell Proliferation; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005123:Eye; D005615:Freezing; D008297:Male; D051379:Mice; D051381:Rats; D013737:Testis / Q000187:drug effects*; D014183:Transplantation, Heterologous
D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
Apoptosis; Cryopreservation; Cryoprotectant; Meiosis; Protein expression; Testis