Vijayakumar R, Simoni J, Ndubisi B, DeLeon F, Heine W
Department of Obstetrics and Gynecology, Texas Tech University Health Sciences Center, School of Medicine, Lubbock 79410.
Arch Androl. 1987;19(2):149-58. doi: 10.3109/01485018708986810.
A total of 2070 two-cell mouse embryos were recovered from 89 superovulated female hybrid mice. Six different culture media were tested. The various media supported mouse embryo development as follows (percentage mean +/- SD, n = 10): Hopp and Pitts medium (H&P) 87 +/- 5 Dulbecco's modified; Eagle's medium supplemented with 10% (volume/volume, v/v) fetal bovine serum (DMEM) 80 +/- 4; Ham's F-10 +/- 15.0% (v/v) human fetal cord serum (hFCS) 79 +/- 3; Whittingham's T-6 medium (WT-6) 60 +/- 4; Ham's F-10 +/- 7.5% (v/v) hFCS 55 +/- 5; Krebs-Ringer low bicarbonate buffer (KRLBB) 42 +/- 6. In H&P, DMEM, WT-6, and Ham's F-10 medium supplemented with hFCS, the pH was maintained within a narrow range of 7.30-7.45 and adequate level of oxygenation was achieved during 72 h in culture. KRLBB had poor buffering capacity and attained ineffective levels of oxygenation during culture. Superior mouse embryo development from two-cells to morulae and hollow blastocysts occurred in H&P, Ham's F-10 + 15% hFCS, and DMEM. Ham's F-10 medium supplemented with hFCS is routinely checked for its ability to support mouse two-cell embryo development to morulae and blastocysts. This is done in conjunction with H&P medium as the control.
从89只超排的雌性杂交小鼠中总共回收了2070个二细胞期小鼠胚胎。测试了六种不同的培养基。各种培养基对小鼠胚胎发育的支持情况如下(平均值±标准差,n = 10):霍普和皮茨培养基(H&P)87±5;添加10%(体积/体积,v/v)胎牛血清的杜尔贝科改良伊格尔培养基(DMEM)80±4;含15.0%(v/v)人胎脐血清(hFCS)的哈姆F-10培养基79±3;惠廷厄姆T-6培养基(WT-6)60±4;含7.5%(v/v)hFCS的哈姆F-10培养基55±5;克雷布斯-林格低碳酸氢盐缓冲液(KRLBB)42±6。在H&P、DMEM、WT-6以及添加hFCS的哈姆F-10培养基中,pH值维持在7.30 - 7.45的狭窄范围内,并且在72小时的培养过程中实现了充足的氧合水平。KRLBB缓冲能力差,在培养过程中氧合水平无效。在H&P、含15% hFCS的哈姆F-10培养基和DMEM中,二细胞期小鼠胚胎发育成桑椹胚和中空囊胚的情况较好。常规检查添加hFCS的哈姆F-10培养基支持小鼠二细胞胚胎发育成桑椹胚和囊胚的能力。这一检查以H&P培养基作为对照同时进行。