人乳腺上皮细胞（HMEC）

Catalogue No.:MZ-1517Product Format: a T25 flaskOrganism: Homo sapiens (human)Complete Growth Medium: See PropagationSource: Organ: breast Tissue: mammary Disease: normalCellType: epithelialAtmosphere: air, 95%; carbon dioxide (CO2), 5% ，37℃Application: Cells and cancer researchPropagation: The base medium forHMECis formulated RPMI-1640 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum toa final concentration of 10%.NOTE: FOR RESEARCH USE ONLY.Components ：Item a T25 flask ManualSpecifications： 2X106 1copySubculturing: Protocol:1.Remove culture medium to a centrifuge tube.2.Briefly rinse theCelllayer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.3.Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope untilCelllayer is dispersed (usually within 1 to 5 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37C to facilitate dispersal.4.Add 6.0 to 8.0ml of complete growth medium and aspirate cells by gently pipetting.5.Transfer theCellsuspension to the centrifuge tube with the medium and cells fromstep 1, and centrifuge at approximately 125 x g for 5 to 10 minutes. Discard the supernatant.6.Resuspend theCellpellet in fresh growth medium. Add appropriate aliquots of theCellsuspension to new culture vessels.7.Incubate cultures at 37C.Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:5 is recommendedMedium Renewal: Every 2 to 3 daysPreservation: Freeze medium: Culture medium,90%; DMSO, 10%Storage temperature: liquid nitrogen vapor phaseTips：1.细胞经过运输后，部分细胞由于温度变化及剧烈碰撞死亡破碎形成碎片，是正常现象。贴壁细胞可以消化，悬浮细胞直接混匀收集细胞，900-1000rpm(约150g)离心3min，弃上清。加5ml PBS重悬细胞，再900-1000rpm(约150g)离心3min，用新鲜的完全培养基重悬接种到新的培养瓶。第二次PBS重悬是为了去除碎片，如果平时碎片比较少，传代时可以省略PBS重悬的步骤；如果碎片很多，建议PBS多洗几次。2.细胞生长不均时，可以将细胞消化吹散后加入新的培养基重新接种或传代。3.细胞生长缓慢时，可以选择提高血清浓度培养（最高不超过20%），也可以根据细胞生长状态，选择传代细胞到新的培养瓶中继续培养。4. 不同细胞贴壁性差异比较大，所以消化时间差别较大，20s-10min均有可能，具体以细胞消化到相互分离但未脱落，并可以轻轻吹下为准，严禁消化到细胞完全漂浮。客户消化过度导致细胞死亡、漂浮、生长缓慢，不提供免费售后服务。5. 干冰发货均为两支，客户先复苏一支，若复苏失败及时联系我方并在我方指导下复苏第二支。6.细胞状态正常时，应尽快冻存细胞保种，冻存后应随机抽取一支检测冻存效果。

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