资料下载
GES-1(人胃粘膜细胞)
阅读:1056 发布时间:2017-7-5提 供 商 | 上海哈灵生物科技有限公司 | 资料大小 | 169.7KB |
---|---|---|---|
资料图片 | 下载次数 | 117次 | |
资料类型 | PDF 文件 | 浏览次数 | 1056次 |
免费下载 | 点击下载 |
GES-1(人胃粘膜细胞)
1.Origin and General Characteristics
Cell Name | GES-1 |
Organism | Homo sapiens, human |
Age |
|
Tissue |
|
Morphology | epithelial |
Growth Properties | adherent |
Descriptions |
|
Biosafety Level |
|
2.Culture Conditions and Handling | |
Complete Growth Medium | RPMI-1640 (150110)+10% FBS (164210-500)+1% P/S (180120) |
| Remove and discard culture medium. Briefly rinse the cell layer with DPBS |
| solution to remove all traces of serum that contains trypsin inhibitor. |
| Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells |
| under an inverted microscope until cell layer is dispersed (usually within 3 |
Subculturing | to 5 minutes). Cells that are difficult to detach may be placed at 37°C to |
| facilitate dispersal. |
| Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently |
| pipetting. Add appropriate aliquots of the cell suspension to new culture |
| vessels. |
Subc*tion Ratio | 1:2-1:4 |
Medium Renewal | every 2 to 3 days |
Cryopreservation | Freeze medium: 50% basal medium+40% FBS+10% DMSO |
Storage temperature: liquid nitrogen vapor phase | |
| |
Culture Conditions | Atmosphere: Air, 95%; CO2, 5% |
Temperature: 37℃ | |
| |
3.Special Features of the Cell Line | |
Tumorigenic |
|
Effects |
|
Receptor Expression |
|
Antigen Expression |
|
Gene Expression |
|
Applications |
|
收到常温细胞后如何处理?
(细胞培养详细操作步骤请参照邮件附件)
- 首先,观察细胞培养瓶是否完好,培养液是否有漏液、浑浊等现象。若有,请拍照,并及时与(所拍照片将作为后续服务依据)。
- 用 75%酒精擦拭细胞培养瓶表面,显微镜下观察细胞状态。因运输问题,部分贴壁细胞会有少量从瓶壁脱落;先不要打开培养瓶盖,将细胞置于细胞培养箱内静置培养 2-4 小时,以便稳定细胞状态。
- 仔细阅读细胞说明书,了解细胞相关信息,如贴壁特性(贴壁/悬浮)、细胞形态、所用基础培养基、血清比例、所需细胞因子、传代比例、换液频率等。
- 静置完成后,取出细胞培养瓶,镜检、拍照,记录细胞状态(所拍照片将作为后续服务依据);建议细胞传代培养后,定期拍照、记录细胞生长状态。
- 贴壁细胞:若细胞生长密度超过 80%,可正常传代;若未超过 80%,移除细胞培养瓶内培养基,预留 5ml 左右继续培养,直至细胞密度达 80%左右再进行传代操作,瓶盖可稍微拧松。
- 悬浮细胞:将细胞培养瓶内液体全部转移至 50ml 无菌离心管内,1200rpm 离心 5min,离心后上清培养基可收集备用,管底细胞沉淀加入 5ml 培养基吹打、重悬。镜检时,若细胞密度超过 80%,可将细胞悬液分至 2 个细胞培养瓶内培养,补加培养基至 5ml;若细胞密度未超过 80%,将细胞悬液移至原瓶继续培养,直至细胞密度达 80%左右时再进行传代操作。