• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.289-296
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• 2014

Human adipose-tissue-derived stromal cells (hADSCs) are abundant in adipose tissue and can differentiate into multi-lineage cell types, including adipocytes, osteoblasts, and chondrocytes. In order to define the optimal harvest site of adipose tissue harvest site, we solated hADSCs from different subcutaneous sites (upper abdomen, lower abdomen, and thigh) and compared their proliferation and potential to differentiate into adipocytes and osteoblasts. In addition, this study examined the effect of phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, on proliferation and differentiation of hADSCs to adipocytes or osteoblasts. hADSCs isolated from different subcutaneous depots have a similar growth rate. Fluorescence-activated cell sorting (FACS) analysis showed that the expression levels of CD73 and CD90 were similar between hADSCs from abdomen and thigh regions. However, the expression of CD105 was lower in hADSCs from the thigh than in those from the abdomen. Although the adipogenic differentiation potential of hADSCs from both tissue regions was similar, the osteogenic differentiation potential of hADSCs from the thigh was greater than that of hADSCs from the abdomen. Phorbol 12-myristate 13-acetate (PMA) treatment increased osteogenic differentiation and suppressed adipogenic differentiation of all hADSCs without affecting their growth rate and the treatment of Go6983, a general inhibitor of protein kinase C (PKC) blocked the PMA effect. These findings indicate that the thigh region might be a suitable source of hADSCs for bone regeneration and that the PKC signaling pathway may be involved in the adipogenic and osteogenic differentiation of hADSCs.

• Archives of Plastic Surgery
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• v.34 no.1
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• pp.1-7
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• 2007

Purpose: Human adipose tissue-derived stromal cells(hADSCs) can be expanded in vitro and induced to differentiate into multiple mesenchymal cell types. In this study we have examined various neuronal phenotypes and gene expression profiles of the hADSCs in the neuronal induction. Methods: The hADSCs were isolated from human adipose tissue and they were characterized by the flow cytometry analysis using CD13, CD29, CD34, CD45, CD49d, CD90, CD105 and HLA-DR cell surface markers. We differentiated the hADSCs into the neuronal lineage by using chemical induction medium and observed the cells with contrast microscopy. The immunocytochemistry and western blotting were performed using the NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III antibodies. Results: The hADSCs were positive for CD13($90.3{\pm}4%$), CD29($98.9{\pm}0.7%$), CD49d($13.6{\pm}6%$), CD90 ($99.4{\pm}0.1%$), CD105($96%{\pm}2.8%$) but negative for CD34, CD45 and HLA-DR. The untreated cultures of hADSCs predominately consisted of spindle shaped cells and a few large, flat cells. Three hours after the addition of induction medium, the hADSCs had changed morphology and adopted neuronal-like phenotypes. The result of immunocytochemistry and western blotting showed that NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III were expressed. However, NSE, NeuN, Vimentin were weakly expressed in the control. Conclusion: Theses results indicate that hADSCs have the capabillity of differentiating into neuronal lineage in a specialized culture medium. hADSCs may be useful in the treatment of a wide variety of neurological disorders.

• Journal of Life Science
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• v.27 no.7
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• pp.767-782
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• 2017

MicroRNAs control the differentiation and proliferation of human adipose tissue-derived stromal cells (hADSCs). However, the role of miR-200a and miR210 on the osteogenic differentiaton of hADSCs has not been determined. hADSCs were isolated from human adipose tissues. Direct binding of mircoRNA to target mRNAs was determined by luciferase assay of the constructs containing putative microRNA binding sites within 3' untranslated region of target mRNAs. Overexpression of miR-200a increased the proliferation and osteogenic differentiation of hADSCs, while causing downregulation of the levels of ZEB2. Inhibition of miR-200a with antisense RNAs inhibited the proliferation and osteogenic differentiation of hADSCs. Overexpression of miR-210 was found to inhibit the proliferation of hADSCs but increase the osteogenic differentiation, while causing downregulation of the levels of IGFBP3. Inhibition of miR-210 with antisense RNAs increased the proliferation but inhibited the osteogenic differentiation of hADSCs. Analysis of the luciferase activity of the constructs containing the miR-200a target site within the ZEB2 3' region and the miR-210 target site within the IGFBP3 3' region revealed lower activity in the miR-200a- or miR-210-transfected hADSCs than in control miRNA-transfected hADSCs. Downregulation of ZEB2 or IGFBP3 in the hADSCs showed similar effects on both their proliferation and osteogenic differentiation with that of miR-200a and miR-210 overexpression, respectively. The results of the current study indicate that miR-200a and miR-210 regulate the osteogenic differentiation and proliferation of hADSCs through the direct targeting of IGFBP3 and ZEB2, respectively.

• Biomolecules & Therapeutics
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• v.17 no.4
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• pp.353-361
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• 2009

Recent in vitro and in vivo animal studies have reported that statin, a cholesterol-lowering drug, stimulate osteogenic differentiation. In the present study, we investigated the effect of simvastatin on osteogenic and adipogenic differentiation in primarily cultured human adipose-derived stem cells (hADSCs). The simvastatin treatment significantly increased the positive cell numbers in alkaline phosphatase and von Kossa staining, and enhanced the expression levels of bone morphogenic protein (BMP)-2, core binding factor alpha 1 (cbfa1), collgen type I and osteonectin mRNAs. Lastly, hADSCs were cultured in the adipogenic media with or without simvastatin to examine the effect of simvastatin on adipogenic differentiation. In the RT-PCR analysis, there were notable decreases in mRNA expression of aP1, C/EBP-$\alpha$ and PPAR-$\gamma$ in hADSCs cultivated in simvastatin-added medium, compared to those in simvastatin-free medium. It suggests that the adipogenic differentiation was significantly inhibited by simvastatin treatment. These observations indicate that simvastatin induces osteogenic differentiation and suppresses adipogenic differentiation in hADSCs.

• Korean Chemical Engineering Research
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• v.51 no.6
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• pp.727-732
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• 2013

In this report, we present an inverse opal scaffold that can enhance the chondrogenic differentiation of human adipose-derived stem cells (hADSCs) without drug, gene, or cytokine supplement. Inverse opal scaffolds based on poly(D,L-lactide-co-glycolide) were formed with uniform $200{\mu}m$ pores. Due to uniform pore sizes and well-controlled interconnectivity of inverse opal scaffold, hADSCs were allowed to distribute homogeneously throughout the scaffolds. As a result, high cell density culture with scaffold was possible. Since the hADSCs cultured in inverse opal scaffolds were subjected to limited supplies of oxygen and nutrients, these cells were naturally preconditioned to a hypoxic environment that stimulated the up-regulation of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). As a result, apoptotic activity of hADSCs until 3 weeks after initial cell seeding was significantly reduced and chondrogenic differentiation related molecular signal cascades were up regulated (transforming growth factor-beta, phosphorylated AKT, and phosphorylated p38 expression). In contrast, hADSCs cultured with small and non-uniform porous scaffolds showed significantly increased apoptotic activity with decreased chondrogenic differentiation. Taken together, inverse opal scaffold could potentially be used as an effective tool for improving chondrogenesis using stem cells.

• Archives of Plastic Surgery
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• v.38 no.2
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• pp.131-134
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• 2011

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 ($20{\mu}M/50{\mu}M\;H_2O_2$), 4) adipogenesis induction culture medium containing ROS ($20{\mu}M/50{\mu}M\;H_2O_2$) and antioxidant ($10{\mu}M/20{\mu}M$ 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 $H_2O_2$ (in a $H_2O_2$ dose-dependently manner) than in media containing adipogenesis induction culture medium and no $H_2O_2$ (p<0.001). Furthermore, in media containing adipogenesis induction culture medium, $H_2O_2$, and antioxidant, absorbance results were significantly lower than in adipogenesis induction culture medium and $H_2O_2$ (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.

• Journal of Life Science
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• v.23 no.4
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• pp.471-478
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• 2013

Human adipose tissue-derived mesenchymal stem cells (hADSCs) have therapeutic potential, including the ability to self-renew and differentiate into multiple lineages. Understanding of molecular mechanisms of stem cell differentiation is important for improving the therapeutic efficacies of stem cell transplantation. In this study, we determined the role of nuclear factor of activated T cells (NFAT5) in the osteogenic differentiation of hADSCs. The down-regulation of NFAT5 expression by the transfection of a specific siRNA significantly inhibited osteogenic differentiation of hADSCs and decreased the activity of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) promoter without affecting their proliferation and adipogenic differentiation. The inhibition of NFAT5 expression inhibited the basal and Tumor Necrosis Factor ${\alpha}$ (TNF-${\alpha}$) induced activation of NF-${\kappa}B$, but it did not affect TNF-${\alpha}$-induced degradation of the $I{\kappa}B$ protein. These findings indicate that NFAT5 plays an important role in the osteogenic differentiation of hADSCs through the modulation of the NF-${\kappa}B$ pathway.

• Biomedical Science Letters
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• v.26 no.4
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• pp.249-255
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• 2020

Radiation therapy is one of the primary options for the treatment of malignant tumors. Even though it is an effective anti-cancer treatment, it can cause serious complications owing to radiation-induced damage to the normal tissue around the tumor. It was recently reported that normal stem cell response to the genotoxic stress of ionizing radiation can boost the therapeutic effectiveness of radiation by repairing damaged cells. Therefore, we focused on annexin A-1 (ANXA1), one of the genes induced by low-dose irradiation, and assessed whether it can protect adipose-derived stem cells (ADSCs) against oxidative stress-induced damage caused by low-dose irradiation and improve effectively cell survival. After confirming ANXA1 expression in ADSCs transfected with an ANXA1 expression vector, exposure to hydrogen peroxide (H2O2) was used to mimic cellular damage induced by a chronic oxidative environment to assess cell survival under oxidative conditions. ANXA1-transfected ADSCs demonstrated that increased viability compared with un-transfected cells and exhibited enhanced anti-oxidative properties. Taken together, these results suggest that ANXA1 could be used as a potential therapeutic target to improve the survival of stem cells after low-dose radiation treatment.

• BMB Reports
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• v.47 no.8
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• pp.469-474
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• 2014

Cell therapies utilizing mesenchymal stem cells (MSCs) have a great potential in many research and clinical settings. The mechanisms underlying the therapeutic effects of MSCs have been studied previously and the paracrine effects elicited by their production of various growth factors and cytokines were recognized as being crucial. However, the molecular controls that govern these paracrine effects remain poorly understood. To elucidate the molecular regulators of this process, we performed a global knockdown of microRNAs (miRNAs) in human adipose-derived mesenchymal stem cells (hADSCs) by inhibiting DGCR8, a key protein in miRNA biogenesis. Global disruption of miRNA biogenesis in hADSCs caused dramatic changes in the expression of subsets of growth factors and cytokines. By performing an extensive bioinformatic analysis, we were able to associate numerous putative miRNAs with these genes. Taken together, our results strongly suggest that miRNAs are essential for the production of growth factors and cytokines in hADSCs.

• Childhood Kidney Diseases
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• v.19 no.2
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• pp.89-97
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• 2015

Background: We conducted this experimental study to examine whether human adipose-derived stem cells (ADSCs) are effective in achieving a recovery of damaged renal tubular epithelial cells in an animal model of cisplatin-induced acute kidney injury using rats. Methods: To examine the in vitro effects of ADSCs in improving nephrotoxicity, we treated mouse renal tubular epithelial cells with both ADSCs and cisplatin mouse renal tubular epithelial cells. And we equally divided 30 male white Sprague-Dawley (SD) rats into the three groups: the control group (intraperitoneal injection of a sterile saline), the cisplatin group (intraperitoneal injection of cisplatin) and the ADSC group (intraperitoneal injection of cisplatin and the hADSC via the caudal vein). At five days after the treatment with cisplatin, serum levels of blood urine nitrogen (BUN) and creatinine were measured from each SD rat. We performed histopathologic examinations of tissue samples obtained from the kidney. Results: The degree of the expression of TNF-${\alpha}$ and that of Bcl-2 were significantly higher and lower respectively, in cisplatin group (P<0.05). Serum levels of BUN (P=0.027) and creatinine (P=0.02) were significantly higher in cisplatin group. On histopathologic examinations, there was a significant difference in the ratio of the renal injury between cisplatin group and ADSC group (P=0.002). Conclusion: The ADSCs might have a beneficial effect in regenerating the damaged renal tubular epithelial cells.