Desiccation tolerance of spermatozoa dried at ambient temperature: production of fetal mice.
Long-term preservation of mouse sperm by desiccation is economically and logistically attractive. The current investigation is a feasibility study of the preservation of mouse sperm by convective drying in an inert gas (nitrogen). Mouse sperm from the B6D2F1 strain isolated in an EGTA-supplemented Tris-HCl buffer were dried using three different drying rates and were stored for 18-24 h at 4 degrees C. The mean final moisture content was <5% for all the protocols. After intracytoplasmic sperm injection (ICSI), the mean blastocyst formation rates were 64%, 58%, and 35% using the rapid-, moderate-, and slow-drying protocols, respectively. The slow-drying protocol resulted in a rate of development significantly lower than that observed using rapid- and moderate-drying protocols and indicated that a slower drying rate may be detrimental to the DNA integrity of mouse sperm. The transfer of 85 two- or four-cell embryos that were produced using rapidly desiccated sperm resulted in 11 fetuses (13%) on Day 15 compared with the production of 34 fetuses (40%) produced using the transfer of 86 two- or four-cell embryos that were produced using fresh sperm (P < 0.05). The results demonstrate the feasibility of using a convective drying protocol for the successful desiccation of mouse sperm and identifies some of the important parameters required for optimization of the procedure.
Bhowmick
Sankha
S
Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston 02115, USA.
Zhu
Liben
L
McGinnis
Lynda
L
Lawitts
Joel
J
Nath
Bharat D
BD
Toner
Mehmet
M
Biggers
John
J
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002021:Buffers; D018481:Convection; D003470:Culture Media; D003890:Desiccation / Q000379:methods*; D004492:Edetic Acid / Q000494:pharmacology; D010064:Embryo Implantation / Q000502:physiology; D005314:Embryonic and Fetal Development / Q000502:physiology; D005260:Female; D005440:Fluid Therapy; D005612:Freeze Drying; D066298:In Vitro Techniques; D007315:Insemination, Artificial / Q000379:methods*; D008297:Male; D051379:Mice; D005741:Noble Gases; D009865:Oocytes / Q000254:growth & development; D010063:Ovum / Q000502:physiology; D011247:Pregnancy; D013094:Spermatozoa / Q000502:physiology*; D013696:Temperature
D002021:Buffers; D003470:Culture Media; D005741:Noble Gases; D004492:Edetic Acid