DMEM高糖培养基(含L-谷氨酰胺,丙酮酸钠110mg/L;不含HEPES,双抗)
DMEM (Dulbecco’s Modified Eagle Medium) is a widely used basal medium for supporting the growth of many different mammalian cells. Cells successfully cultured in DMEM include primary fibroblasts, neurons, glial cells, HUVECs, and smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12. We offer a variety of DMEM modifications for a range of cell culture applications..
This DMEM is modified as follows:
储存条件:2-8° C
运输条件:常温
使用建议:DMEM is unique from other media as it contains 4 times the concentration of amino acids and vitamins than the original Eagle’s Minimal Essential Medium. DMEM was originally formulated with low glucose (1 g/L) and sodium pyruvate, but is often used with higher glucose levels, with or without sodium pyruvate. DMEM contains no proteins, lipids, or growth factors. Therefore, DMEM requires supplementation, commonly with 10% (FBS). DMEM uses a sodium bicarbonate buffer system (3.7 g/L), and therefore requires a 5–10% CO2 environment to maintain physiological pH.
质控指标及依据
- 外观及澄清度(中华人民共和国药典2010版二部附录ⅨB)
- pH值(中华人民共和国药典2010版二部附录ⅥH)
- 渗透压(中华人民共和国药典2010版二部附录ⅨG)
- 细菌内毒素(中华人民共和国药典2010版二部附录ⅨE)
- 微生物限度(中华人民共和国药典2010版二部附录ⅨJ)
- 细胞生长试验(中华人民共和国化工行业标准《哺乳类动物细胞培养基》 HG/T3935-2007第707条)
细胞生长试验
检测依据:中华人民共和国化工行业标准《哺乳类动物细胞培养基》 HG/T3935-2007第707条
方法:
- 以5×10^4/ml细胞数将细胞接种到6孔板中,每种细胞每种培养基6个孔(48h检测用3个孔+48h换液用3个孔),起始培养基为含有5%FBS的待测培养基。
- 培养箱中静置培养48h后,将每个板中3个检测孔的细胞消化收集,并使用细胞计数仪进行计数和存活率测定。
- 将3个换液孔中的培养基吸弃,并加入无FBS的待测培养基。
- 继续在培养箱中静置培养48h后,将换液孔中细胞消化,使用细胞计数仪进行计数和存活率测定。
稳定性实验
- 检测依据:行业标准及ICH指导原则
- 常规稳定性数据:推荐保质期为12个月,分别在2-8度储存条件下,检测3个月,6个月,12个月,16个月稳定性,结果无明显差异,符合质控标准
加速破坏实验:将产品存放于电子恒温箱内,分别测试在37摄氏度,45摄氏度,条件下的稳定性,每个温度下,分为三组,正置,倒置,水平放置,选取8小时,24小时,48小时,72小时节点进行测试,产品稳定性良好,无明显统计学差异
恒温运输模拟测试:将产品在7月12日从大连发往海南,普通非冷藏包装,申通快递运输,于7月19日到达,然后由对方立即申通快递返回,于7月27日收到,然后进行检测,产品稳定性良好,无统计学差异,符合恒温运输条件
培养基对比同类进口产品结果展示:
培养基名称 | 细胞 | 品牌 | +5%FBS 培养48h | 换液无FBS 培养48h | ||
细胞数(个/ml) | 存活率 | 细胞数(个/ml) | 存活率 | |||
DMEM高糖 | Vero | 进口品牌 | 1.60×10^5 | 92.1% | 3.86×10^5 | 92.6% |
美仑MA0212 | 1.65×10^5↑ | 93.8% | 3.70×10^5 | 95.6% | ||
AGS | 进口品牌 | 1.24×10^5 | 89.7% | 1.60×10^5 | 86.3% | |
美仑MA0212 | 1.39×10^5 ↑ | 88.9% | 1.92×10^5 ↑ | 90.4% | ||
293T | 进口品牌 | 1.06×10^5 | 85.0% | 4.36×10^5 | 90.2% | |
美仑MA0212 | 1.08×10^5 ↑ | 85.2% | 4.45×10^5 ↑ | 90.1% | ||
L-929 | 进口品牌 | 1.80×10^5 | 91.7% | 2.29×10^5 | 93.9% | |
美仑MA0212 | 1.77×10^5 | 94.1% | 2.59×10^5 ↑ | 94.3% | ||
L-929 | 进口品牌 | 1.06×10^5 | 94.3% | 1.19×10^5 | 82.8% | |
美仑MA0212 | 1.09×10^5 ↑ | 95.8% | 1.18×10^5 | 95.0% | ||
293 | 进口品牌 | 3.15×10^5 | 91.3% | 3.73×10^5 | 89.1% | |
美仑MA0212 | 3.43×10^5 ↑ | 93.6% | 4.07×10^5 ↑ | 89.8% | ||
Vero | 进口品牌 | 1.16×10^5 | 91.2% | 2.25×10^5 | 92.9% | |
美仑MA0212 | 1.21×10^5 ↑ | 93.0% | 2.77×10^5 ↑ | 94.8% | ||
RAW 264.7 | 进口品牌 | 2.02×10^6 | 90.3% | 1.01×10^6 | 74.9% | |
美仑MA0212 | 2.65×10^6 ↑ | 89.7% | 1.57×10^6 ↑ | 83.8% |
结论:美仑基础培养基在培养细胞的细胞数和存活率方面均与进口品牌相媲美,且显微镜下细胞形态良好,长期培养细胞效果稳定,可以完美取代进口培养基。
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