CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells.
CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells.
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DOI：
10.1186/bcr1371
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Smalley MJ
中科院分区：
文献类型：
--
作者：
Sleeman KE;Kendrick H;Ashworth A;Isacke CM;Smalley MJ
乳腺癌被认为是由乳腺上皮干细胞引起的。因此，对识别这些细胞有很大的兴趣。乳房是由两个上皮层（外部肌上皮/基底层和内腔上皮层）以及大的非上皮组分（成纤维细胞、内皮细胞、淋巴细胞、脂肪细胞、神经元和肌细胞）组成的复杂组织。乳腺上皮干细胞群的确定性鉴定关键取决于其纯度。迄今为止，这一直受到缺乏合适的标记物来分离两个上皮层和去除污染的非上皮细胞的阻碍。分离小鼠乳腺并用CD 24染色。基于CD 24表达将细胞分选成单独的群体，并通过直接荧光显微镜和真实的PCR评估腔上皮和肌上皮/基底标志物。这些细胞群的干/祖细胞潜能通过清除的乳腺脂肪垫移植在体内进行评估。鉴定了三种表达CD 24的细胞群：CD 24阴性、CD 24低和CD 24高。用细胞角蛋白标记物对这些细胞进行染色显示，这些群体分别对应于非上皮、肌上皮/基底和腔上皮细胞。通过定量PCR确认细胞身份。这些细胞群的清除的乳腺脂肪垫移植揭示了广泛的乳腺脂肪垫再增殖能力与CD 24低细胞分离，而CD 24高细胞具有有限的再增殖能力。乳腺上皮细胞对CD 24的差异染色可用于同时分离非上皮、肌上皮/基底和腔上皮细胞的纯群体。此外，乳腺脂肪垫再增殖能力在CD 24低群体中富集。由于使用单一标记物实现分离，因此将可能并入额外的标记物以进一步细分这些群体。这将大大促进乳腺上皮细胞亚群的进一步分析，同时确保高纯度，这是了解正常组织生物学和致癌作用中乳腺上皮干细胞的关键。
Breast cancer is thought to arise in mammary epithelial stem cells. There is, therefore, a large amount of interest in identifying these cells. The breast is a complex tissue consisting of two epithelial layers (an outer myoepithelial/basal layer and an inner luminal epithelial layer) as well as a large non-epithelial component (fibroblasts, endothelial cells, lymphocytes, adipocytes, neurons and myocytes). The definitive identification of a mammary epithelial stem cell population is critically dependent on its purity. To date, this has been hampered by the lack of suitable markers to separate out the two epithelial layers, and to remove contaminating non-epithelial cells. Mouse mammary glands were dissociated and stained with CD24. Cells were sorted into separate populations based on CD24 expression and assessed for luminal epithelial and myoepithelial/basal markers by direct fluorescent microscopy and real time PCR. The stem/progenitor potential of these cell populations was assessed in vivo by cleared mammary fat pad transplantation. Three populations of CD24 expressing cells were identified: CD24Negative, CD24Low and CD24High. Staining of these cells with cytokeratin markers revealed that these populations correspond to non-epithelial, myoepithelial/basal and luminal epithelial cells, respectively. Cell identities were confirmed by quantitative PCR. Cleared mammary fat pad transplantation of these cell populations revealed that extensive mammary fat pad repopulation capacity segregates with the CD24Low cells, whilst CD24High cells have limited repopulation capacity. Differential staining of mammary epithelial cells for CD24 can be used to simultaneously isolate pure populations of non-epithelial, myoepithelial/basal and luminal epithelial cells. Furthermore, mammary fat pad repopulation capacity is enriched in the CD24Low population. As separation is achieved using a single marker, it will be possible to incorporate additional markers to further subdivide these populations. This will considerably facilitate the further analysis of mammary epithelial subpopulations, whilst ensuring high purity, which is key for understanding mammary epithelial stem cells in normal tissue biology and carcinogenesis.
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影响因子：
2.5
作者：
Smalley, MJ;Clarke, RB
通讯作者：
Clarke, RB
DOI：
10.1073/pnas.0502320102
发表时间：
2005-05-10
影响因子：
11.1
作者：
Xin, L;Lawson, DA;Witte, ON
通讯作者：
Witte, ON
影响因子：
3.8
作者：
Smith, GH
通讯作者：
Smith, GH
影响因子：
2.7
作者：
Clarke, RB;Spence, K;Potten, CS
通讯作者：
Potten, CS
DOI：
10.1084/jem.183.4.1797
发表时间：
1996-04-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Goodell MA;Brose K;Paradis G;Conner AS;Mulligan RC
通讯作者：
Mulligan RC