种属：人

性别，年龄：女，36岁

组织来源：乳腺

生长特性：贴壁生长形态特征：上皮细胞样

细胞特性：The MCF 10A cell line is a non-tumorigenic epithelial cell line.

The cells are positive for epithelial sialomucins, cytokeratins and milk fat globule antigen.

They exhibit three dimensional growth in collagen, and form domes in confluent cultures.

Thus far, the cells have shown no signs of terminal differentiation or senescence.

The line is responsive to insulin, glucocorticoids, cholera enterotoxin, and epidermal growth factor (EGF).

By electron microscopy the cells display characteristics of luminal ductal cells but not of myoepithelial cells.

They also express breast specific antigens as detected by positive reaction with MFA-Breast and MC-5 monoclonal antibodies.

The calcium content of the medium exerts a strong effect on the morphology of the cells.

支原体及其它微生物检测：阴性

安全性：所有肿瘤和病毒转染的细胞均视为有潜在的生物危害性，必须在二级生物安全台内操作，并请注意防护，所有废液及接触过此细胞的器皿需高压灭菌后方能丢弃。

培养条件：

完全培养基：The base medium for this cell line (MEBM) along with the additives can be obtained from Lonza/Clonetics Corporation as a kit: MEGM, Kit Catalog No. CC-3150. ATCC does not use the GA-1000 (gentamycin-amphotericin B mix) provided with kit. To make the complete growth medium, you will need to add the following components to the kit (sold separately):

100 ng/ml cholera toxin

Note: Do not filter complete medium

或A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium, 20 ng/ml Human epidermal growth factor, 100 ng/ml cholera toxin, 0.01 mg/ml bovine insulin and 500 ng/ml hydrocortisone, 95%; horse serum, 5%

培养温度：37 ℃；CO2：5%。

传代方法：

收到细胞后，取出培养瓶在倒置显微镜下观察细胞生长情况：

如果细胞未长满，用75%酒精喷洒整个瓶后放到超菌台内，严格无菌操作，打开细胞培养瓶，吸出培养液，加新的 10ml培养液在瓶内继续培养。

如果细胞已长满，即可进行传代培养。具体步骤如下：

1. 弃去培养液，用PBS(不含钙，镁离子)洗1-2次。

2. 加0.5ml消化液(0.125%Trypsin-0.25mM EDTA)于培养瓶中，倒转放于37度培养箱1-3分钟预热，然后又将培养瓶倒转过来大约30秒后，在倒置显微镜下观察细胞消化情况，若细胞大部分变圆分散，轻敲几下培养培养瓶，细胞随即脱落下来。

3. 加入6-8mlDMEM+１０%ＦＢＳ培养液或１-２ｍｌ胰酶抑制剂，吸入离心管，离心(１０００转，6分钟)，弃上清，加１０ｍｌ完全培养液将细胞重悬，分到新的培养瓶中。一传二或三。

注：1、观察细胞**在低倍镜(4或5X物镜)下进行，否则不能准确判断细胞的传代密度。

2、瓶中运输培养基不能重复再用,请换用加双抗的新培养基，细胞冻存后,培养基中可不加任何抗生素。

3、有些细胞贴壁不牢，如发现贴壁细胞有脱落，可离心吹打后接种到新瓶内。

4、收到细胞后，若发现培养瓶破损、有液溢出及细胞有污染，请及时与我们联系。

冻存方法：

冻存液：90%完全培养液，10%DMSO。现用现配

储存：液氮储存