Testicle sperm freeze-storage and revival method
A sperm and testicular technology, applied in the field of testicular sperm cryopreservation and recovery, can solve the problems of low motility, low testicular sperm count, and failure of cryopreserved testicular sperm recovery, etc., and achieves the effect of improving flexibility and efficiency.
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Embodiment 1
[0013] Put the seminiferous tubules taken out from the biopsy into a small amount of modified human fallopian tube fluid (mHTF), crush the seminiferous tubules with elbow forceps, blow and beat them evenly with a pipette, and prepare testicular sperm by discontinuous density gradient separation method, and put them into 10% polyvinylpyrrolidone (PVP) microdroplets in intracytoplasmic sperm injection (ICSI) dishes.
[0014] The zona pellucida was derived from donated eggs in first meiotic prophase, metaphase or unfertilized eggs. The zona pellucida is prepared by using a micromanipulation system on an inverted microscope hot stage. The egg cells are fixed with an egg-holding needle with an inner diameter of 15um, and a hole is made at the 3 o'clock position with an injection needle with an inner diameter of 5um, and the cytoplasm is aspirated with the injection needle , until the zona pellucida becomes empty. The zona pellucida stays in the modified human fallopian tube fluid ...
Embodiment 2
[0018] Referring to the method of Example 1, a total of 34 zona pellucida are used to freeze sperm, of which 8 zona pellucida freeze testicular sperm, as a reference, the other 12 zona pellucida freeze few weak sperm, and 14 zona pellucida freeze normal sperm . A total of 530 live sperm were injected into the zona pellucida. Before freezing, each zona pellucida contained 15.6 sperm on average. After recovery, an average of 12.3 live sperm and 1.8 dead sperm were recovered from each zona pellucida. The recovery rate of testicular sperm was 71.7%. The sperm recovery rate was 78.7%. See Table 1 for the results.
[0019] sperm type
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