Characterization in vitro of luminal and myoepithelial cells isolated from the human mammary gland by cell sorting

O’Hare, Michael J.; Ormerod, M.G.; Monaghan, Paul; Lane, E. Birgitte; Gusterson, Barry A.

doi:10.1111/j.1432-0436.1991.tb00883.x

Cited by 114 publications

(88 citation statements)

References 35 publications

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“…Further characterization by FACS analysis of specific surface antigens showed EMA positivity on about 50% of epithelial cells. As predicted, there was no evidence for CALLA, a myoepithelial cell marker (O'Hare et al, 1991), in these cells.…”

Section: Genetic Analysissupporting

confidence: 64%

Short-term primary culture of epithelial cells derived from human breast tumours

Speirs

Green²,

Walton³

et al. 1998

Br J Cancer

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Summary As experimental models for breast cancer, most studies rely on established human breast cancer cell lines. However, many of these lines were established over 20 years ago, many from pleural effusions rather than the primary tumour, so the validity of using them as representative models is questionable. This paper describes our experiences, over a 3-year period, in establishing short-term epithelial-cellenriched preparations from primary breast tumours based on differential centrifugation followed by culture in selective media. Epithelial cells were successfully cultured from 55% of samples, but culture success did not appear to be correlated with tumour histology, stage, grade or node status. Epithelial cell-enriched cultures were immunopositive for broad-spectrum cytokeratin and epithelial membrane antigen (EMA). Positivity for keratin 19 confirmed that the cultures contained tumour-derived cells, which additionally showed significantly higher activity of the reductive pathway of the steroid-converting enzyme 17p-hydroxysteroid dehydrogenase type 1. That the cultures contained tumour and not normal epithelial cells was further substantiated by the complete absence of the calmodulin-like gene NB-1 in tumour-derived cultures; this is only associated with normal breast epithelia. Eighty-five per cent of cultures established from oestrogen receptor (ER)-positive tumours expressed ER in vitro; this was functional in 66% of cultures, although ER-positive phenotype was gradually lost over time. In conclusion, epithelial cells can be isolated and maintained as short-term cultures from primary breast tumours irrespective of histopathological or clinical details, providing a model system with a greater biological and clinical relevance than breast cancer cell lines.

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Section: Genetic Analysissupporting

confidence: 64%

Short-term primary culture of epithelial cells derived from human breast tumours

Speirs

Green²,

Walton³

et al. 1998

Br J Cancer

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“…The concept of markers for normality can be referred to as the "canary in the coal mine" principle whereby the muting of a singing canary at the bottom of a coal mine signals a poisonous gas leak early enough to allow immediate evacuation of miners. 101 Examples of this principle in diagnostic pathology include a variety of basal cell markers (eg, p63, K5, K14, SMA), the presence of which argues strongly against malignancy in a variety of lesions in the breast, 102,103 salivary glands, 104,105 and prostate. 106 -111 Although gastric chief cells are distinct from basal or myoepithelial cells (not present in gastrointestinal epithelium), MIST1 expression allows determination of an intact chief cell compartment on an individual gland basis.…”

Section: Discussionmentioning

confidence: 99%

The Transcription Factor MIST1 Is a Novel Human Gastric Chief Cell Marker Whose Expression Is Lost in Metaplasia, Dysplasia, and Carcinoma

Lennerz

Kim

Oates

et al. 2010

The American Journal of Pathology

105

148

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The lack of reliable molecular markers for normal differentiated epithelial cells limits understanding of human gastric carcinogenesis. Recognized precursor lesions for gastric adenocarcinoma are intestinal metaplasia and spasmolytic polypeptide expressing metaplasia (SPEM), defined here by ectopic CDX2 and TFF2 expression, respectively. In mice, expression of the bHLH transcription factor MIST1, normally restricted to mature chief cells, is down-regulated as chief cells undergo experimentally induced metaplasia. Here, we show MIST1 expression is also a specific marker of human chief cells. The mainstays of therapy in gastric carcinoma are early recognition, resection, and neoadjuvant or adjuvant therapy. However, gastric cancer remains the second largest cause of cancer-related mortality worldwide, 1 which drastically illustrates our lack of understanding of the sequence and progression of preneoplastic conditions. The traditional linear progression model of cellular changes, such as (Helicobacter-mediated) inflammation, atrophy, intestinal metaplasia (IM), dysplasia, and carcinoma, 2-4 does not apply to all cases and does not allow incorporation of more recently recognized entities. For example, there are distinct types of IMs, not all carrying definitive preneoplastic potential, and some authors have argued that IM in general is a paraneoplastic condition because the earliest gastric carcinomas arise from gasSupported by

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“…In a similar manner, stem cells were shown to reside in proximal ducts of the prostate [25]. In their original paper, DeOme et al [1] observed that any portion of the mouse mammary gland contained cells that could give rise to another and growth, and a similar conclusion was drawn also for human breast [26]. However, through careful microscope-directed collection of mammary organoids from reduction mammoplasty followed by multiple culture assays we recently demonstrated that the terminal ducts are one of the major repositories of stem cells in adults [19].…”

Section: Identifying Human Mammary Stem Cells By Their Nichementioning

confidence: 53%

Of Microenvironments and Mammary Stem Cells

LaBarge

Petersen²,

Bissell

2007

Stem Cell Rev

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In most adult tissues there reside pools of stem and progenitor cells inside specialized microenvironments referred to as niches. The niche protects the stem cells from inappropriate expansion and directs their critical functions. Thus guided, stem cells are able to maintain tissue homeostasis throughout the ebb and flow of metabolic and physical demands encountered over a lifetime. Indeed, a pool of stem cells maintains mammary gland structure throughout development, and responds to the physiological demands associated with pregnancy. This review discusses how stem cells were identified in both human and mouse mammary glands; each requiring different techniques that were determined by differing biological needs and ethical constraints. These studies together create a robust portrait of mammary gland biology and identify the location of the stem cell niche, elucidate a developmental hierarchy, and suggest how the niche might be manipulated for therapeutic benefit. KeywordsNiche; Stem cells; Mammary gland; Microenvironment Making A Case for A Mammary Stem CellThe mammary gland is a hormone sensitive, branching, and bilayered epithelial organ; it consists of an inner luminal epithelial layer and an outer myoepithelial layer surrounded by a basement membrane in a stromal fat pad. Cells from the two epidermal layers express a number of different proteins that can be used to unambiguously identify them. Among the most common of these markers are keratin (K) 8, K18, K19, and sialomucin-1 (Muc-1) expressed by luminal cell; and K5, K14, and α-smooth muscle actin expressed by myoepithelial cells. The arbor-like gland is organized into lobules that are inter-connected by a system of ducts; the basic functional unit of the breast is termed the terminal ductal lobular unit (TDLU). The gland is considered to have differentiated functionally and completely during lactation when the alveoli are swollen, producing milk, and secreting it into the lumen. Because the mammary gland has the ability to undergo repeated cycles of expansion (as much as tenfold at pregnancy) followed by involution, back to a state nearly indistinguishable from the virgin glands, it is clear that it is endowed with cells with remarkable regenerative capacity.

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Characterization in vitro of luminal and myoepithelial cells isolated from the human mammary gland by cell sorting

Cited by 114 publications

References 35 publications

Short-term primary culture of epithelial cells derived from human breast tumours

Short-term primary culture of epithelial cells derived from human breast tumours

The Transcription Factor MIST1 Is a Novel Human Gastric Chief Cell Marker Whose Expression Is Lost in Metaplasia, Dysplasia, and Carcinoma

Of Microenvironments and Mammary Stem Cells

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