Shackleton M, Vaillant F, Simpson KJ et al (2006) Generation of a functional mammary gland from a single stem cell. J Intern Med 264:128–142Ĭao Y, Sun Z, Liao L et al (2005) Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Morani A, Warner M, Gustafsson JA (2008) Biological functions and clinical implications of oestrogen receptors alfa and beta in epithelial tissues. Nature 465:798–802Ĭheng G, Weihua Z, Warner M et al (2004) Estrogen receptors ER alpha and ER beta in proliferation in the rodent mammary gland. Breast Cancer Res 8:R49Īsselin-Labat ML, Vaillant F, Sheridan JM et al (2010) Control of mammary stem cell function by steroid hormone signalling. Dev Biol 277:443–456īooth BW, Smith GH (2006) Estrogen receptoralpha and progesterone receptor are expressed in label-retaining mammary epithelial cells that divide asymmetrically and retain their template DNA strands. Recent Prog Horm Res 14:219–248 discussion 248–254Ĭlarke RB, Spence K, Anderson E et al (2005) A putative human breast stem cell population is enriched for steroid receptor-positive cells. Lyons WR, Li CH, Johnson RE (1958) The hormonal control of mammary growth and lactation. Dev Biol 325:106–121īai L, Rohrschneider LR (2010) s-SHIP promoter expression marks activated stem cells in developing mouse mammary tissue. Kurpios NA, MacNeil L, Shepherd TG et al (2009) The Pea3 Ets transcription factor regulates differentiation of multipotent progenitor cells during mammary gland development. Woodward WA, Chen MS, Behbod F et al (2005) On mammary stem cells. Smalley MJ (2010) Isolation, culture and analysis of mouse mammary epithelial cells. Sleeman KE, Kendrick H, Ashworth A et al (2006) CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells. Stingl J, Eirew P, Ricketson I et al (2006) Purification and unique properties of mammary epithelial stem cells. Welm BE, Tepera SB, Venezia T et al (2002) Sca-1(pos) cells in the mouse mammary gland represent an enriched progenitor cell population. Our research demonstrates that Sca-1 + mammary stem cells can be more easily isolated when cultured in the presence of oestrogen and GH.Īl-Hajj M, MF Clarke (2004) Self-renewal and solid tumor stem cells. Furthermore, gene expression analysis by Real-time PCR revealed that Sca-1 + cells expressed markedly higher levels of mammary stem cell-related genes in comparison to Sca-1 − cells. We confirmed that Sca-1 + cells regenerated mammary outgrowths when transplanted in vivo, formed mammospheres in vitro and differentiated into luminal epithelial cells with milk-secreting function and myoepithelial cells under Matrigel culture. Flow cytometry showed that 50% of cells were Sca-1 + under the culture of MaECM medium. We established MaECM medium supplemented with oestrogen and growth hormone (GH), in which oestrogen promoted mammary epithelial cell proliferation and inhibited fibroblast growth, and GH obviously enhanced the effect of oestrogen on mammary epithelial cell proliferation. Stem cell medium BM was used to culture mammary organoids, which generated many fibroblasts. We first selected the medium suitable for mammary stem cell growth. Real-time PCR was carried out to analyse the expression of mammary stem cell-related genes in Sca-1+ cells. Differentiating culture was used to detect the differentiation potential of Sca-1 + cells. Mammary fat pad transplantation and Mammosphere-forming assay were done to confirm the stem cell potential of Sca-1 + cells. Flow cytometry was used to isolate Sca-1 + and Sca-1 − cell populations from cultured mammary epithelial cells. We used BM medium supplemented with different concentration of 17 MaECM medium which promoted the proliferation of mouse mammary epithelial cells and inhibited the growth of fibroblasts. The object of our study is to establish the Medium to promote the proliferation of mammary stem cell and explore the possibility of Sca-1 as mammary stem cell marker. Sca-1 is controversial as a mammary stem cell marker in the literature, which may be due to the different isolation protocols and culture media used in different laboratories.
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