Cell & Molecular Biology
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/30432
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Browsing Cell & Molecular Biology by Author "Al-Ghadban, Sara"
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Item Characterization of adipose tissue and adipose-tissue derived stem cells in Lipedema(2021) Al-Ghadban, Sara; Bunnell, BrucePurpose: Lipedema is a painful adipose tissue (AT) disorder that occurs almost exclusively in women, with onset manifesting at puberty or at times of hormonal change. This disorder is characterized by a symmetrical increase of fat deposition in the legs and the arms, sparing the hands and the feet. The goal of this study was to determine the histology of the skin and fat of lipedema tissue and characterize the adipose tissue-derived stem cells (ASCs) obtained from the stromal vascular fraction (SVF) of thigh AT of non-lipedema and lipedema patients. Methods: Histological sections from AT were stained with H&E. Adipocyte area was quantified using ImageJ software. Markers for macrophages (CD68) was investigated by immunohistochemistry. ASCs were characterized by the expression of stemness markers and their multi-differentiation potential. Flow cytometry, RT-PCR and immunofluorescence assays were performed. Results: The data show hypertrophic adipocytes, increased numbers of macrophages and blood vessels in thigh tissue of women with lipedema compared to non-lipedema patients. Additionally, at the cellular level, an increase in the adipogenic differentiation potential with no change in the expression of mesenchymal markers (CD73, CD90 and CD105) or extracellular markers (collagen, fibronectin and laminin) was detected in lipedema ASCs compared to non-lipedema ASCs. Conclusion: Infiltration of immune cells, increase in adipocyte size and adipogenesis stimulates angiogenesis and fibrosis in lipedema AT. Defining the structure and the components of AT will provide insights into the pathophysiology of lipedema and will help researchers develop potential treatment for the disease.Item Molecular characterization of adipose-tissue derived stem cells (ASCs) from the breast(2021) Artiles, Maria; Bunnell, Bruce; Al-Ghadban, SaraPurpose: It has been reported that both the health of the donor, as well as the site of collection can alter the proliferation and differentiation properties of adipose tissue-derived stem cells (ASCs) . Obesity alters ASCs and induces them to enhance the tumorigenic properties of breast cancer cell lines both in vitro and in vivo. Abdominal ASCs (aASCs) from obese donors show increased expression of leptin and estrogen. Leptin signaling cascades and estrogen-mediated pathways can increase breast cancer proliferation, tumorigenesis and metastasis. The aim of this study is to characterize ASCs derived from breast tissue (bASCs) of human donors and compare their proliferation and differentiation properties to those of aASCs, and to understand if bASCs are affected in the same way. Methods: bASCs will be characterized and compared to aASCs. Adipogenic and osteogenic differentiation will be induced and assessed with Oil Red O and Alizarin Red S staining, respectively. Colony forming capabilities will be assessed with crystal violet staining. Surface markers will be evaluated with flow cytometry. Expression of adipogenic genes will be assessed by real-time PCR. Results: bASCs are expected to display CD90, CD105 and CD73 surface markers. They are also expected to successfully undergo adipogenic and osteogenic differentiation and to be capable to form colonies. Finally, it is expected to observe differences in proliferation rates between aASCs and bASCs. Conclusion: Characterizing bASCs, and comparing them to aASCs, will provide us an understanding of the unique properties of this subpopulation and their potential applications.