PMC

Effects of CPMs on post-thaw total and progressive motility of C57BL/6J sperm preserved in cryovials. Experiments were repeated 3 times using sperm pooled from 4~6 males.

The cooling curve of a sperm sample in a cryovial with 100 µl sperm in R18S3+MTG submerged in LN vapor for 10 min.

LN Dewar setup for freezing. A, the depth of LN at the Dewar bottom, 2.0~2.5cm; B, the distance between cryovial bottom and LN surface, 0.5~1.0cm; C, the distance between cryovial bottom and the opening of the Dewar, 11 cm. The Dewar was covered with a lid during sperm cooling.

Changes in progressive motility of C57BL/6J sperm during incubation in 4 CPMs at ~23°C. Experiments were repeated 3 times using sperm pooled from 4 to 6 males each experiment.

Changes in total motility of C57BL/6J sperm during incubation in 4 CPMs at ~23°C. Experiments were repeated 3 times using sperm pooled from 4 to 6 males each experiment.

Comparisons of GSH IVF rates of C57BL/6J sperm cryopreserved in cryovials with 50 µl sperm each in R18S3, R18S3+MTG and RSGlu87. The mean of each IVF rate is shown on the top of each bar. Experiments were repeated 3 times using sperm pooled from 4~6 males.

Comparisons of GSH IVF rates of C57BL/6J sperm cryopreserved in cryovials with 150, 100 and 50 µl sperm each and cryostraws with 50, 25 and 10 µl sperm each. R18S3+MTG was used as CPM, and experiments were repeated 3 times using sperm pooled from 4 to 6 males.

IVF rates of frozen-thawed sperm of C57BL/6J (n=3), C57BL/6N (n=3), GM lines on C57BL6J background (n=9), and GM lines on C57BL/6N background (n=54), compared with that of fresh C57BL/6J sperm (n=3). B6J = C57BL/6J; B6N = C57BL/6N; BKGD = background; n =number of sperm cryopreservation (SC) procedures = number of IVF procedures. Each SC or IVF procedure used sperm from 1 male in GM mouse lines or pooled from 4 to 6 males in B6J and B6N wild-type strains. Sperm were collected in R18S3+MTG (0.5 ml per male) and cryopreserved in cryovials with 50 µl sperm.