McGinnis Lynda K, Zhu Liben, Lawitts Joel A, Bhowmick Sankha, Toner Mehmet, Biggers John D
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Biol Reprod. 2005 Oct;73(4):627-33. doi: 10.1095/biolreprod.105.042291. Epub 2005 Jun 1.
Mouse sperm with and without trehalose were desiccated under nitrogen gas and stored at 4 degrees C and 22 degrees C. After rehydration, sperm were injected into oocytes using intracytoplasmic sperm injection and embryonic development was followed. Sperm were dried for 5.0, 6.25, or 7.5 min, stored at 22 degrees C for 1 wk with and without trehalose. The percentages of blastocysts that developed from sperm with trehalose were 51%, 31%, and 20%, respectively, which was significantly higher than sperm without trehalose (10%, 3%, and 5%, respectively). Desiccation and storage in medium with trehalose significantly increased sperm developmental potential compared to medium without trehalose. Sperm dried for 5 min produced more blastocysts than sperm dried for 6.25 or 7.5 min. When sperm were dried in trehalose for 5 min and stored for 1 wk, 2 wk, 1 mo, or 3 mo at 4 degrees C, the percentages of blastocysts were 73%, 84%, 63%, and 39%; whereas those stored at 22 degrees C for 1 wk, 2 wk, or 1 mo were significantly lower (53%, 17%, and 6%, respectively). Embryos from sperm partially desiccated in trehalose for 5 min and stored at 4 degrees C for 1 or 3 mo were transferred to 10 pseudopregnant recipients. Implantation rates were 81% and 48%; live fetuses were 26% and 5%, respectively. One of the recipients delivered three live fetuses. The results show that trehalose has a significant beneficial effect in preserving the developmental potential of mouse sperm following partial desiccation and storage at temperatures above freezing.
将含有和不含海藻糖的小鼠精子在氮气下干燥，并分别储存在4℃和22℃。复水后，使用胞浆内单精子注射将精子注入卵母细胞，并跟踪胚胎发育情况。精子干燥5.0、6.25或7.5分钟，在有和没有海藻糖的情况下于22℃储存1周。含有海藻糖的精子发育成囊胚的比例分别为51%、31%和20%，显著高于不含海藻糖的精子（分别为10%、3%和5%）。与不含海藻糖的培养基相比，在含有海藻糖的培养基中进行干燥和储存可显著提高精子的发育潜能。干燥5分钟的精子产生的囊胚比干燥6.25或7.5分钟的精子更多。当精子在海藻糖中干燥5分钟，并在4℃下储存1周、2周、1个月或3个月时，囊胚的比例分别为73%、84%、63%和39%；而在22℃下储存1周、2周或1个月的囊胚比例则显著较低（分别为53%、17%和6%）。将在海藻糖中部分干燥5分钟并在4℃下储存1或3个月的精子所形成的胚胎移植到10只假孕受体体内。着床率分别为81%和48%；活胎率分别为26%和5%。其中一只受体产下了三只活胎。结果表明，海藻糖在保存部分干燥并在高于冰点的温度下储存的小鼠精子的发育潜能方面具有显著的有益作用。
