Cryopreservation method for trace sperms

• Main Authors: FENG CHUN, LYU PINGPING, CHEN XIJING, QIAN YULI, YE YINGHUI, LI JINGPING
• Format: Patent
• Language: Chinese; English
• Published: 24.10.2023
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS; PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTSTHEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TRAPPING; VINEGAR; WINE

The invention discloses a cryopreservation method of trace sperms, which comprises the following steps: (1) pre-treating a sperm sample to prepare a sperm suspension; (2) uniformly mixing the sperm suspension with a sperm cryopreservation solution according to a volume ratio of 1: 1 to obtain a uniformly mixed sperm suspension; (3) sequentially sucking samples from the open end of the straw according to the following sequence: a. 80-150mu L of mHTF culture medium containing 10% of serum; b, air with the length of 15-25 mm; c. 5-10 [mu] L of a uniformly mixed sperm suspension; d, air with the length of 15-25 mm is adopted; (4) sealing the open end of the straw; and the sealed straw is cryopreserved by liquid nitrogen. The straw made of plastic is not easy to break in the freezing and unfreezing process and can be heat-sealed, so that the hidden danger of liquid nitrogen pollution is reduced; each straw can be frozen to be as low as 5 microliters of sperm suspension, so that the volume is reduced, the cooling a