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Immunohistochemical Study of CD24 in Mammary Epithelial Small Light Cells during Different Development Stage

Author: ZengXiaoFang
Tutor: AnZuo
School: Southern Medical University,
Course: Medical Developmental Biology
Keywords: Breast Stem cell Morphology Immunohistochemistry
CLC: S852.4
Type: Master's thesis
Year: 2008
Downloads: 68
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Abstract

The rodents of mammary epithelial First appear in the breast with a future glandular epithelial buds form. 3 weeks after stimulation by sex hormones, duct system into the fat pad and formation of branches, known as the terminal buds, the starting point for the growth of mammary duct, the cell mass is wrapped around the body of the layer by the goblet cells. With the development of catheter stretching and terminal buds, somatic form the catheter internal cavity epithelium and goblet cells forming the muscle on the external cortex. The catheter lumen is formed in the process, the body cells will usually undergo apoptosis, and goblet cells also into a body cell layer, so that the goblet cells are sources of stem cells. The mature breast there may be cells having stem cell function. In rodents, pregnancy, rapid cell proliferation, leading to a large number of branch like a bunch of grapes and secretory vesicles form will appear. In secretory vesicles, the the luminal epithelium milk, to myoepithelial contraction milk secretory vesicles to squeeze into the catheter. Once weaned, these structures disappear by apoptosis, like a non-pregnant, the conduit structure of the fat pad also become quite sparse. This cycle is gone through a number of animal life. The the only proliferative potential of stem cells can maintain the process; suggests the presence of temporary stem cells, subjected to sexual stimulation, these cells in a quiescent state. Moreover, the the incomplete process also seen in the estrous cycle. The same cycle is also found in human pregnancy and the menstrual cycle, but there are certain differences in the proliferation extent. In addition, the day-to-day breast cell renewal process, the the mature mammary stem cells (mammary stemcells, MaSCs) may also play a role. Shedding of epithelial cells in the breast-feeding process, milk. Falling into the cells of the conduit system in some way must be supplemented, or the number of epithelial cells can not maintain integrity; cells having such a function may be provided having a continuous active stem cells. So mature mammary epithelial stem cells having at least two distinctly different functions, i.e. temporary or persistent stem cells. Although there are stem cells in the breast tissue, but there is still a lack of specific markers of breast stem cells, many scholars have applied, including breast ball cultivate 5 - bromo-2 - deoxy - uridine (BrdU) labeling method, cell surface markers such as stem cell antigen 1 (Sca-1) and-CD49f, Hoechst staining a series of methods to identify breast stem cells. Especially based on the surface antigen MaSCs phenotype analysis is the study of stem cell biology characteristics and effectively select MaSCs, clinical transplantation in the treatment of a key technology, the MaSCs surface antigen analysis technology has been widely used for the separation of cell subsets, which Sca- 1 molecule is the most common the application of MaSCs selection marker. But in recent years, information has confirmed The Sca-1 ^{- -MaSCs of existence, Stingl such as the use of flow cytometry sorted CD49f ^{/ Sca-1 ^{hi View mammary epithelial cells in transplantation experiments and communities, CD49f ^{/ Sca-1 ^{low in clean fat pad there strong the reconstruction ability possessed-CD49f ^{phenotype of mammary epithelial cells, After training will eventually express Sca-1, for the Scal ^{- -MaSCs than Scal ^{-MaSCs. Alvi of mammary side population cells (hematopoietic stem / progenitor cells isolated with Hoechst dye and flow cytometry, a group of special cells called side population cells, it has a similar stem cell self-renewal and differentiation potential, that is a universal stem cell phenotype.) found, the SP and the non-SP have similar colony forming ability, despite its contained ^{-Sca-1 the proportion of cells was significantly different (15.8% Sca-1 in SP and1.8% Sca-1 in non-SP cells). Therefore, it is necessary to find a new, more specific stem cell surface markers. A new antigen CD24 cause for concern, recently Shackleton breast tissue into single cell suspension screened with special surface markers by flow cytometry cells and injected LacZ transgenic mice, and found that all inoculated CD29 ^{hi Lin ^{- of CD24 ^{cells of mice at 10 weeks, a significant clone. Although the research is not very in-depth, but its application are enticing. , There is a form of special epithelial cells in the mouse and rat mammary ducts and acinar. Light microscope, this smaller cell bodies were round nucleus strong basophilic round centered nucleoplasm cytoplasm containing organelles less the cytoplasm transparent, clear, and is known as the small bright cells, similar to bright cells are also found in the human breast. Now widely recognized that this small bright cells may represent a MaSCs. Adult rat model using histology, immunohistochemistry chemical and light microscopy detection technology adult rat mammary tissue of the development of the law, as well as the surrounding tissue, the occurrence of evolution; adult rat mammary tissue CD24-positive small bright cells express volume and expression site changes, comparisons between the line Kruskal-Wallis test method for group, pairwise comparisons Nemenyi law line group, take the test level α = 0.05. The various parts of the experimental results are as follows: Part I: morphological study of the different developmental stages of the adult rat mammary HE staining was observed 3-month-old is not pregnant, pregnant 13 days, seven days postpartum, four periods of 21 days postpartum rat mammary tissue. The experimental results show that, not pregnant 3 months and 21 days postpartum breast tissue is mostly connective tissue and fat tissue, epithelial ductal and acinar small, isolated or in clusters scattered in fat or connective tissue. Located in acinar or catheter, there is a small bright cells smaller cell bodies, they are distributed in the ductal and acinar basement membrane or myoepithelial glandular epithelial cells. Them with clear boundaries of adjacent cells, and no exposure to the lumen or acinar cavity surface. 13 and pregnant, postpartum 7 days mammary epithelial cells proliferate extensively in the milk ducts matrix acinar developmental increase in the number of larger gland epithelial cell body, can be more easily observed small bright cells in ductal and acinar can even find cell mass by 1-3 small bright cells close to the basement membrane. Part II: the adult rat mammary gland at different developmental stages small bright cells CD24 immunohistochemical study and statistical analysis of immunohistochemical observation selected four periods (nonpregnant age of 3 months and 13 days pregnant, postpartum 7 days 21 days postpartum, respectively, corresponding to the stationary phase of the breast tissue, breast tissue, pregnancy, lactation, degradation period), in terms of the catheter or acinar epithelium are visible CD24-positive small bright cells, and in the catheter the degree of staining is generally higher than the acini. Statistical analysis of the results (A: not pregnant, 3 months old, B: 13 days of pregnancy, C: postpartum 7 days, D: 21 days postpartum): ① the Kruskal-Wallis method, χ ^{2 = 21.36, P < 0.005; ② Nemenyi method, X ^{2 _{A, B} = 12.02, P <0.05; X ^{2 _{A, C,} = 12.53, P <0.05; X ^{2 _{A, D} = 0.15, P> 0.05; X ^{2 _{B, C} = 0.03, P> 0.05; X ^{2 _{B, D} = 8.79, P <0.05; X ^{2 _{C, D} = 9.29, P <0.05. Pregnancy and lactation compared to the stationary phase and the degradation period, CD24 expressed more small bright cells during pregnancy and lactation; quiescent and degradation period showed no significant difference (P> 0.05); expression was also no significant difference (P> 0.05).}}}}}}}}}}}}}}}}}}}

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