藏酋猴精液深低温冻存的研究──不同的冷冻程序、冷冻稀释保存液冻存效果的比较

SEMEN CRYOPRESERVATION IN THE TIBETAN MACAQUE (MACACA THIBETANA)——COMPARISON OF DIFFERENT COOLING PROGRAMS AND FREEZING MEDIA

以存活率（ＳＲ）、复苏运动度（ＲＭ）、ＳＰＡ为精子活力检测指标，对四种冷冻程序ＰＳＦ、Ｈ３Ｐ、ＬＰ、ＭＤＰ和卵黄的、无卵黄的冷冻稀释保存液进行了藏酋猴（Ｍａｃａｃａ ｔｈｉｂｅｔａｎａ）精液冻存比较研究。 ＰＳＦ的ＳＲ为９０．１±１．９％，ＲＭ为６３．８±２．８％，显著高于其他三种冷冻程序（Ｐ＜０．０１）。卵黄冷冻稀释保存液（ＭＤＭ）的ＳＲ（９５． ４±１．３％）和ＲＭ（８８．０±１０．２％）显著高于无卵黄防冻液（ＦＣＳ－Ｇ／ＴＨ）的ＳＲ（９０．１±１．９％）和ＲＭ（６３．６±２．８％），但前者在复苏一小时后，ＲＭ（１２．５±１．６％）明显低于后者的ＲＭ（５４． ７±２． ２％）。用 ＦＣＳ～Ｇ／ＴＨ冻存的精液经 ＳＰＡ检测，其穿透率为新鲜精子的 ５１． ９％。结果提示：１）慢速降温冷冻程序适于藏酋猴精液冻存；２）卵黄冷冻稀释保存液能较好地保存藏酋猴精液的活动度；而无卵黄防冻液则有利于延长其冷冻精子的运动寿命。这可能与两者的稀释液及所含的脂蛋白不同有关。

Four cooling programmes (PSF,H3P,LP and MDP) and four freezing media,including three egg-yolk media (MDM,DM,PM)and one non-egg-yolk medium (FCS-G/TH),were compared in cryopreservation of the Tibetan macaque semen. Of the four cooling programmes,the best results which were achieved by PSF,featured slow cooling rate and automatic ice seeding. Following the program PSF, the sperm survival rate (SR)and recovery motility (RM)were 90. 1±1. 9%and 63.8 ± 2.8%, respectively, which were superior to those of LP(SR, 73.71 ±2.0%;RM,42.4±2.5% ), H3P(SR, 66.3±1.0% ; RM,38.7±1.8% ),MDP(SR, 35.8±1. 6%;RM, 15.3±1.0%) (P<O.01). The SRs and RMs were negatively related to the cooling rates of the four procedure used. of the three egg-yolk media MDM,DM and PM,the RMs were 88.0±10.2 %, 46.2 ±3.2 %, 43.2±2.4% ,respectively. MDM which had low concentration of egg yolk and glycerol,gave significantly higher RM (p< 0.01). In comparison between MDM and FCS-G/TH, MDM had significantly higher (p<0.05) sperm survival rate (SR, 95.4± 1.2 % )and recovery motility (RM, 88.0±10.2 % ) than that following FCS-G/TH (SR, 90.1±1.9%; RM, 63.6±2.8 % ). One hour after thawing, however, MDM' S recovery motility dropped dramatically to 12.5±2.3 %,whereas FCS-G/TH's recovery motility remained as high as 54.7±3.0%. Fertilization viability of FCS-G/TH frozen sperm was tested in sperm penetration assay (SPA)and the penetration rate of the store frozen sperm was 51.9% of that freshly collected sperm. These results suggested that: 1 ). Cryopreservation protocol constitute with cooling pro-gramme of slow cooling rate,ice seeding (PSF),freezing media of low concentration of egg yolk and glycerol (e. g. MDM, FCS-G/TH)benefits semen cryopreservation in the Tibetan macaque; 2). Egg yolk medium (MDM)keeps much more motility,but non-egg yolk medium (FCS-G / TH ) preserve longer life span of motile sperm. This might be due to the different composition of two freezing media.