Effect of EGF on angiogenesis induced by adipose-derived stem cells / 中华医学美学美容杂志

Objective To explore the effect of epidermal growth factor (EGF) on tube formation of HUVEC induced by the secretion of angiogenesis factors of adipose-derived stem cells (ADSCs).Methods ADSCs were primarily cultured by enzyme digestion method.The flow cytomertry was performed to detect the expression of cell surface marker.ELISA was used to detect the expression of VEGF,HGF,and SDF-1 after given different doses of EGF.Tube formation assay was used to examine the effect of EGF on the tube formation induced by ADSCs.Results ADSCs were successfully isolated and cultured from human liposuction tissue and specific markers were expressed on ADSCs.EGF promoted the secretion of angiogenesis factors VEGF,HGF,and SDF-1,which were secreted by ADSCs.EGF pretreatment increased the ability of tube formation of HUVECs induced by ADSCs.Conclusions ADSCs induce the secretion of angiogenesis factors in vitro,and thus increase the ability of tube formation of HUVECs.EGF promotes the secretion ability of ADSCs,and the best concentration is 15 mg/L.

Effect of platelet-rich fibrin on proliferation and adipogenic differentiation of adipose-derived stem cells / 中华医学美学美容杂志

Objective To study the effect of autogeneic platelet-rich fibrin (PRF) on proliferation and adipogenic differentiation of human adipose-derived stem cells (ADSCs) in vitro.Methods ADSCs were isolated from adipose tissue obtained from donors undergoing liposuction and were cultured,and underwent identification.ADSCs at passage 3 were divided into three groups:test groups were cultured with 1PRFM and 2PRFM,and control group was cultured without PRF membrane.Then the growth of the cells was observed by inverted microscope.MTT method was used to observe cell proliferation activity at days 1,2,3,4,5,6 and 7 after culture.Adipogenic differentiation of ADSCs was observed and quantified by oil red O staining at days 3,5,7,9,11 and 14.Results Cell proliferation and adipogenic differentiation would be increased with the PRFM,There were significant differences among three groups.Conclusions PRF could significantly promote proliferation and adipogenic differentiation of ADSCs.

Role of Reactive Oxygen Species in the Adipogenesis of Adipose-derived Stem Cells

PURPOSE: Stem cells continue to receive research attention in the clinical fields, and adipose-derived stem cells(ADSCs) have been shown to be a good source raw material. Many plastic surgeons are researching the ADSC adipogenesis with a view of conducting clinical trials, and many attempts have been made to identify the factors that promote the adipogenesis of ADSCs, but comparatively few correlation studies have been undertaken to explore the relation between reactive oxygen species(ROS) and the ADSC adipogenesis. We undertook this study is to investigate the effects of ROS on ADSC adipogenesis. METHODS: ADSCs were isolated and cultured from abdominal adipose tissue, and cultured in different media; 1) DMEM(control), 2) adipogenesis induction culture medium, 3) adipogenesis induction culture medium with ROS(20 microM/50 microM H2O2), 4) adipogenesis induction culture medium containing ROS(20 microM/50 microM H2O2) and antioxidant(10 microM/20 microM Deferoxamine). We compared adipogenesis in these different media by taking absorbance measurements after Oil-Red O staining every 5 days. RESULTS: After culturing for 20 days, significant differences were observed between these various culture groups. Absorbance results showed significantly more adipogenesis had occurred in media containing adipogenesis induction culture medium and H2O2(in a H2O2 dose-dependently manner) than in media containing adipogenesis induction culture medium and no H2O2(p<0.001). Furthermore, in media containing adipogenesis induction culture medium, H2O2, and antioxidant, absorbance results were significantly lower than in adipogenesis induction culture medium and H2O2(p<0.001). CONCLUSION: These findings suggest that ROS promote the adipogenesis of ADSCs. We suggests that ROS could be used in the adipose tissue engineering to improve fat cell differentiation and implantable fat tissue organization.

Characterization and differentiation into adipocytes of mesenchymal stem cells (MSCs) from human adipose tissue and amniotic fluid / 대한산부인과학회지

OBJECTIVE: Mesenchymal stem cells (MSCs) are potentially very useful for regenerative and reparative medicine as well as therapeutic possibilities. The aim of this study is to examine the ability of ADSCs and AFSCs to be phenotypically and functionally differentiated into adipocyte and to determine the appropriate stem cell source and conditions for efficient adipocyte regeneration. METHODS: Adipogenic differentiation was induced by culturing confluent ADSCs and AFSCs in adipogenic medium for 2~4 weeks. During the differentiation inducing period, we evaluated the successful adipogenesis by performing immunocytochemistry and RT-PCR to detect the lipid producibility and several adipogenic gene expressions including lipoprotein lipase (LPL), peroxisome proliferator-activated receptor gamma2 (PPAR gamma2) and adiponectin. RESULTS: ADSCs and AFSCs are expanded easily in vitro and exhibited a fibroblast-like morphology as previously known in MSCs from bone marrow and a commercial source. Flow cytometric analysis showed that ADSCs and AFSCs expressed several CD marker antigens similar to those observed on bone marrow-derived MSCs. Adipogenic induction of ADSCs and AFSCs resulted in the extended cell morphology, intracellular staining of an established lipid dye Oil Red O, and expression of adipocyte-specific genes. CONCLUSION: Both ADSCs and AFSCs successfully differentiate in vitro into adipogenic cells in the presence of the lineage-specific induction factors although ADSC showed the greater capability. Therefore, the results suggest that ADSCs and AFSCs may be an excellent choice for many future tissue engineering strategies and cell-based therapies.

The effect of long-term culture in vitro on biological characteristics of human adipose tissue derived stem cells / 中华创伤骨科杂志

Objective To observe the effect of lnng-term in vitro culture on the biological properties of adipose-derived stem cells(ADSCs)as seeding cells of tissue engineering.Methods The surface makers and apoptosis of primary and passaged human ADSCs were identified by flow cytometric analysis.Osteogenic differentiation of ADSCs at different passages were identified by alkaline phosphatase(ALP),Von Kossa staining and RT-PCR respectively.Results The surface marker expression of mesenchymal stem cells on ADSCs was high and did not change with passages of the cells.The early apoptosis rate of the cells was 1% to 2%,and increased insignificantly from passage one to passage nine.The osteogenic potential of ADSCs confirmed by ALP,Von Kossa staining and RT-PCR was maintained to as late as passage eight.Conclusion Since the biological properties of ADSCs are stable,they can be served as optimal seeding cells for tissue engineering and regenerative research.