Harvesting, Isolation and Differentiation of Rat Adipose-Derived Stem Cells

Megaloikonomos, Panayiotis D.; Panagopoulos, Georgios N.; Bami, Myrto; Igoumenou, Vasileios G.; Dimopoulos, Leonidas; Milonaki, Adamantia; Kyriakidou, Maria; Mitsiokapa, Evanthia; Anastassopoulou, J.; Mavrogenis, Andreas F.

doi:10.2174/1389201019666180418101323

Cited by 33 publications

(19 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rat adipose-derived stem cells were harvested via surgery and enzymatically digested from the inguinal fat pads of female Lewis rats as previously described [4 , 5] . Cells were cultured in flasks in a humidified 37 °C, 5% CO 2 incubator.…”

Section: Experimental Design Materials and Methodsmentioning

confidence: 99%

Cytotoxic and apoptotic data of BPA and BPA alternatives TMBPF, BPAF, and BPS in female adult rat and human stem cells

2021

View full text Add to dashboard Cite

Here, we used female adult rat adipose-derived stem cells (rASCs) and human adipose-derived stem cells (hASCs) to compare the toxicities and potencies of several widespread environmental toxins that may be endocrine-disrupting chemicals, including bisphenol A (BPA), and the newer BPA alternatives bisphenol S (BPS), bisphenol AF (BPAF), and tetramethyl bisphenol F (TMBPF). Adult stem cells were cultured to 80% confluency in vitro and then exposed to BPA (1 and 10 µM), 17β-estradiol (E2; 10 µM), BPS (1 and 10 µM), BPAF (3 × 10 −3 –30 µM), TMBPF (0.01–50 µM), or control media alone (with 0.01% ethanol) for varying time intervals from 20 min to 5 hrs. Using several cellular assays, the levels of cell death, apoptosis, caspase-6 activation, and potencies were compared across chemical treatments and vehicle-treated controls. There was significantly decreased cell viability and increased apoptosis in rat and human stem cells treated with each BPA analog, as early as 20 min of exposure, and at low doses. With higher magnification, higher resolution imaging it was evident that in many of the BPA analog-treated cells, the Apopxin Deep Red dye indicative of apoptosis was localized to the cytoplasmic compartments of cells, while the nuclear green DCS1 dye indicative of late-stage apoptosis and necrosis was localized to the nuclei of cells. Notably, BPAF and TMBPF showed cytotoxicity in a dose-dependent manner (BPAF LC 50 = 0.014 µM (rASCs) and 0.009 µM (hASCs); TMBPF LC 50 = 0.88 µM (rASCs) and 0.06 µM (hASCs); lethal concentration with 50% survival). The rank order of potency was BPAF>TMBPF>BPA>BPS. The majority of cell death was due to apoptosis as indicated by high levels of activated caspase-6 in the cytoplasm of almost 100% of cells treated with the BPA analogs. This data allows for further confirmation of caspase-6-mediated apoptosis using higher magnification imaging that definitively demonstrate the cytotoxic and apoptotic effects of these BPA analogs. For a complete description, interpretation, and discussion of the data refer to the article in press [1] .

show abstract

Section: Experimental Design Materials and Methodsmentioning

confidence: 99%

Cytotoxic and apoptotic data of BPA and BPA alternatives TMBPF, BPAF, and BPS in female adult rat and human stem cells

2021

View full text Add to dashboard Cite

show abstract

“…Porcine adipose tissue-derived stem cells (PrADSCs) were isolated from fat surgically extracted from the neck area of experimental pigs (breed Prestice black pied pigs with a weight of approximately 35-40 kg; Institute of Animal Science, Přeštice, Czech Republic) during the surgery under general anesthesia. The protocol for cell isolation was also set according to Estes et al [49] with some modifications developed in studies focused on adipose-derived stem or stromal cells [74,75] and is described in our previous study by Matejka et al 2020 [70]. The cells were expanded in a standard way until the 2 nd passage in DMEM-F12K (Sigma-Aldrich, St. Louis, MO, USA) medium (ratio 1:1) supplemented with 10 % of FBS (Sigma-Aldrich, St. Louis, USA), 1 % of ABAM (Antibiotic Antimycotic Solution, contains 100 units penicillin, 0.1 mg streptomycin, and 0.25 μg amphotericin B per mL of culture media, Sigma-Aldrich, St. Louis, MO, USA) and 10 ng/mL of FGF-2 (GenScript, Piscataway, NJ, USA, Cat.…”

Section: Cell Modelsmentioning

confidence: 99%

Growth Factors VEGF-A165 and FGF-2 as Multifunctional Biomolecules Governing Cell Adhesion and Proliferation

Sedlář

Trávníčková

Matějka

et al. 2021

IJMS

View full text Add to dashboard Cite

Vascular endothelial growth factor-A165 (VEGF-A165) and fibroblast growth factor-2 (FGF-2) are currently used for the functionalization of biomaterials designed for tissue engineering. We have developed a new simple method for heterologous expression and purification of VEGF-A165 and FGF-2 in the yeast expression system of Pichia pastoris. The biological activity of the growth factors was assessed in cultures of human and porcine adipose tissue-derived stem cells (ADSCs) and human umbilical vein endothelial cells (HUVECs). When added into the culture medium, VEGF-A165 stimulated proliferation only in HUVECs, while FGF-2 stimulated the proliferation of both cell types. A similar effect was achieved when the growth factors were pre-adsorbed to polystyrene wells. The effect of our recombinant growth factors was slightly lower than that of commercially available factors, which was attributed to the presence of some impurities. The stimulatory effect of the VEGF-A165 on cell adhesion was rather weak, especially in ADSCs. FGF-2 was a potent stimulator of the adhesion of ADSCs but had no to negative effect on the adhesion of HUVECs. In sum, FGF-2 and VEGF-A165 have diverse effects on the behavior of different cell types, which maybe utilized in tissue engineering.

show abstract

“…ADSCs are multipotent cells with the potential to differentiate into multilineage cells including osteocytes, myocytes, chondrocytes, adipocytes, astrocytes, and endotheliocytes in vitro and in vivo [ 39 ]. However, significantly fewer stem cells exist in pathologic AT compared to normal AT.…”

Section: Pathological Changes During Lymphedemamentioning

confidence: 99%

Adipose-derived stem cell therapy shows promising results for secondary lymphedema

Pan

2020

WJSC

View full text Add to dashboard Cite

Lymphedema is mainly identified by progressive soft tissue swelling in impaired lymphatic system. Secondary lymphedema attributed to cancer therapy, parasite infection, and trauma remains a serious global disease. Patients with lymphedema suffer swelling, pain, and fatigue, with the dysfunction of the deformed extremities reducing the quality of life and increasing the risk of infection and lymphangiosarcoma. Adipose-derived stem cells (ADSCs) possess prominent regenerative potential to differentiate into multilineage cells, and produce various lymphangiogenic factors, making ADSC therapy a promising approach for lymphedema. The development of lymphedema consists of local inflammation, the fibrosis of lymphatic vessels, and the deposition of adipose fat. Existing animal models do not mimic the chronic inflammation environment, therefore suitable models are required in further studies. Some signal pathways and molecular mechanisms in physiological and pathological lymphagiogenesis remain unclear. In previous animal and human trials, ADSC therapy reduced edema in varying degrees. A larger number of trials with larger samples and longer follow-up periods are required to verify the efficiency and feasibility of ADSC therapy. ADSCs are of easy availability and immune exemption, making them a candidate for lymphedema treatment. Whether ADSCs enhance malignant characteristics or trigger the malignant change deserves further exploration and study before ADSC therapy can be made widely available.

show abstract

Harvesting, Isolation and Differentiation of Rat Adipose-Derived Stem Cells

Cited by 33 publications

References 0 publications

Cytotoxic and apoptotic data of BPA and BPA alternatives TMBPF, BPAF, and BPS in female adult rat and human stem cells

Cytotoxic and apoptotic data of BPA and BPA alternatives TMBPF, BPAF, and BPS in female adult rat and human stem cells

Growth Factors VEGF-A165 and FGF-2 as Multifunctional Biomolecules Governing Cell Adhesion and Proliferation

Adipose-derived stem cell therapy shows promising results for secondary lymphedema

Contact Info

Product

Resources

About