Wu J J, Hu T J, Guo B, Yue Z P, Yang Z T, Zhang X M
Department of Laboratory Medicine, Jilin University, Changchun, China.
Cryo Letters. 2011 Sep-Oct;32(5):402-9.
To develop a procedure for cryopreservation of adult bovine testis tissue, the effects of dimethyl sulphoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), and their concentrations (v/v), as well as different thawing temperatures, on the cell viability of bovine testis tissue after freezing/thawing were examined. The highest testicular cell viabilities came from the media containing DMSO (85.3 ± 1.2 percent), PG (82 ± 1.0 percent) and EG (83.4 ± 1.0 percent) at 10 percent concentration respectively. Using 10 percent DMSO gave significantly higher spermatogonia percentage (61.1 ± 1.2 percent, P < 0.001) than processing with 10 percent PG (54.3 ± 0.6 percent) or 10 percent EG (55 ± 1.8 percent) after differential plating. Thawing in water bath of 37 or 97-100 degree C also provided significantly higher viabilities (85.1 ± 1.0, 85 ± 1.0 percent, P < 0.01, respectively) and spermatogonia percentages (56.6 ± 2.0, 56.6 ± 2.6 percent, P < 0.01, respectively) than that thawing at 4C (23.4 ± 0.8 percent for total viability, 8.97 ± 1.0 percent for spermatogonia percentage). Collectively, 10 percent DMSO and thawing in 37-100 degree C water baths were appropriate for the cryopreservation of bovine testicular tissue and subsequent spermatogonia enrichment.
为开发一种成年牛睾丸组织的冷冻保存方法，研究了二甲基亚砜（DMSO）、丙二醇（PG）、乙二醇（EG）及其浓度（体积/体积），以及不同解冻温度对冷冻/解冻后牛睾丸组织细胞活力的影响。睾丸细胞活力最高的分别是浓度为10%的含DMSO（85.3±1.2%）、PG（82±1.0%）和EG（83.4±1.0%）的培养基。使用10%的DMSO进行差异铺板后，精原细胞百分比（61.1±1.2%，P<0.001）显著高于使用10%的PG（54.3±0.6%）或10%的EG（55±1.8%）。在37℃或97 - 100℃水浴中解冻也能提供显著更高的活力（分别为85.1±1.0%、85±1.0%，P<0.01）和精原细胞百分比（分别为56.6±2.0%、56.6±2.6%，P<0.01），高于在4℃解冻（总活力为23.4±0.8%，精原细胞百分比为8.97±1.0%）。总体而言，10%的DMSO和在37 - 100℃水浴中解冻适用于牛睾丸组织的冷冻保存及随后的精原细胞富集。
