• Archives of Plastic Surgery
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• 제35권6호
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• pp.653-658
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• 2008

Purpose: Skin and soft tissue defect is one of the major challenges faced by plastic surgeons. Adipose derived stromal cells, which can be harvested in large quantities with low morbidity, display multilineage mesodermal potential. Therefore, adipose derived stromal cells have been met with a great deal of excitement by the field of tissue engineering. Recently, Adipose derived stromal cells have been isolated and cultured to use soft tissue restoration. In order to apply cultured cells for clinical purpose, however, FDA approved facilities and techniques are required, which may be difficult for a clinician who cultures cells in a laboratory dedicated to research to utilize this treatment for patients. In addition, long culture period is needed. Fortunately, adipose derived stromal cells are easy to obtain in large quantities without cell culture. The purpose of this study is to present a possibility of using uncultured adipose derived stromal cells for wound coverage. Methods: Seven patients who needed skin and soft tissue restoration were included. Five patients had diabetic foot ulcers, 1 patient got thumb amputation, and 1 patient had tissue defect caused by resection of squamous cell carcinoma. The patients' abdominal adipose tissues were obtained by liposuction. The samples were digested with type I collagenase and centrifuged to obtain adipose derived stromal cells. The isolated adipose derived stromal cells were applied over the wounds immediately after the wound debridement. Fibrin was used as adipose derived stromal cells carrier. Occlusive dressing was applied with films and foams and the wounds were kept moist until complete healing. Results: One hundred to one hundred sixty thousand adipose derived stromal cells were isolated per ml aspirated adipose tissue. All patients' wounds were successfully covered with the grafted adipose derived stromal cells in a 17 to 27 day period. No adverse events related to this treatment occurred. Conclusion: The use of uncultured adipose derived stromal cells was found to be safe and effective treatment for wound coverage without donor site morbidity.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.327-333
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• 2006

Future cell-based therapies such as tissue engineering will benefit from a source of autogenous pluripotent stem cells. There are embryonic stem cells (ESC) and autologous adult stem cells, two general types of stem cells potentilally useful for these applications. But practical use of ESC is limited due to potential problems of cell regulation and ethical considerations. To get bone marrow stem cells is relatively burden to patients because of pain, anesthesia requirement. The ideal stem cells are required of such as the following advantages: easy to obtain, minimal patient discomfort and a capability of yielding enough cell numbers. Adipose autologus tissue taken from intraoral fatty pad or abdomen may represent such a source. Our study designed to demonstrate the ability of human adipose tissue-derived stromal cells (hATSC) from human abdominal adipose tissue diffentiating into osteocyte and adipocyte under culture in vitro conditions. As a result of experiment, we identified stromal cell derived adipose tissue has the multilineage potentiality under appropriate culture conditions. And the adipose stromal cells expressed several mesenchymal stem cell related antigen (CD29, CD44) reactions. Secondary, we compared the culture results of a group of hATSC stimulated with TGF-${\beta}$1, bFGF with a hATSC group without growth factors to confirm whether cytokines have a important role of the proliferation in osteogenic differentiation. The role of cytokines such as TGF-${\beta}$1, bFGF increased hATSC's osteogenic differentiation especially when TGF-${\beta}$1 and bFGF were used together. These results suggest that adipose stromal cells with growth factors could be efficiently available for cell-based bone regeneration.

• Archives of Plastic Surgery
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• 제38권4호
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• pp.438-444
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• 2011

Purpose: Adipose-derived stromal cells (ASCs) are readily harvested from lipoaspirated tissue or subcutaneous adipose tissue fragments. The stromal vascular fraction (SVF) is a heterogeneous set of cell populations that surround and support adipose tissue, which includes the stromal cells, ASCs, that have the ability to differentiate into cells of several lineages and contains cells from the microvasculature. The mechanisms that drive the ASCs into the osteoblast lineage are still not clear, but the process has been more extensively studied in bone marrow stromal cells. The purpose of this study was to investigate the osteogenic capacity of adipose derived SVF cells and evaluate bone formation following implantation of SVF cells into the bone defect of human phalanx. Methods: Case 1 a 43-year-old male was wounded while using a press machine. After first operation, segmental bone defects of the left 3rd and 4th middle phalanx occurred. At first we injected the SVF cells combined with demineralized bone matrix (DBM) to defected 4th middle phalangeal bone lesion. We used P (L/DL)LA [Poly (70L-lactide-co-30DL-lactide) Co Polymer P (L/DL)LA] as a scaffold. Next, we implanted the SVF cells combined with DBM to repair left 3rd middle phalangeal bone defect in sequence. Case 2 was a 25-year-old man with crushing hand injury. Three months after the previous surgery, we implanted the SVF cells combined with DBM to restore right 3rd middle phalangeal bone defect by syringe injection. Radiographic images were taken at follow-up hospital visits and evaluated radiographically by means of computerized analysis of digital images. Results: The phalangeal bone defect was treated with autologous SVF cells isolated and applied in a single operative procedure in combination with DBM. The SVF cells were supported in place with mechanical fixation with a resorbable macroporous sheets acting as a soft tissue barrier. The radiographic appearance of the defect revealed a restoration to average bone density and stable position of pharyngeal bone. Densitometric evaluations for digital X-ray revealed improved bone densities in two cases with pharyngeal bone defects, that is, 65.2% for 4th finger of the case 1, 60.5% for 3rd finger of the case 1 and 60.1% for the case 2. Conclusion: This study demonstrated that adipose derived stromal vascular fraction cells have osteogenic potential in two clinical case studies. Thus, these reports show that cells from the SVF cells have potential in many areas of clinical cell therapy and regenerative medicine, albeit a lot of work is yet to be done.

• Journal of Korean Neurosurgical Society
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• 제42권2호
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• pp.118-124
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• 2007

Objective : Adipose tissue is derived from the embryonic mesoderm and contains a heterogenous stromal cell population. Authors have tried to verify the characteristics of stem cell of adipose derived stromal cells (ADSCs) and to investigate immunohistochemical findings after transplantation of ADSC into rat brain to evaluate survival, migration and differentiation of transplanted stromal cells. Methods : First ADSCs were isolated from human adipose tissue and induced adipose, osseous and neuronal differentiation under appropriate culture condition in vitro and examined phenotypes profile of human ADSCs in undifferentiated states using flow cytometry and immunohistochemical study. Human ADSCs were transplanted into the healthy rat brain to investigate survival, migration and differentiation after 4 weeks. Results : From human adipose tissue, adipose stem cells were harvested and subcultured for several times. The cultured ADSCs were differentiated into adipocytes, osteoctye and neuron-like cell under conditioned media. Flow cytometric analysis of undifferentiated ADSCs revealed that ADSCs were positive for CD29, CD44 and negative for CD34, CD45, CD117 and HLA-DR. Transplanted human ADSCs were found mainly in cortex adjacent to injection site and migrated from injection site at a distance of at least 1 mm along the cortex and corpus callosum. A few transplanted cells have differentiated into neuron and astrocyte. Conclusion : ADSCs were differentiated into multilineage cell lines through transdifferentiation. ADSCs were survived and migrated in xenograft without immunosuppression. Based on this data, ADSCs may be potential source of stem cells for many human disease including neurologic disorder.

• 대한두개안면성형외과학회지
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• 제21권4호
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• pp.211-218
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• 2020

Autologous fat grafts are widely used in soft-tissue augmentation and reconstruction. To reduce the unpredictability of fat grafts and to improve their long-term survival, cell-assisted lipotransfer (CAL) was introduced. In this alternative method, autologous fat is mixed and grafted with stromal vascular fraction cells or adipose-derived stem/stromal cells (ASCs). In regenerative medicine, ASCs exhibit excellent therapeutic potential and are also simple to harvest. Although the efficacy of CAL has been demonstrated in experimental and clinical research, studies on its safety in terms of oncologic risk have reported inconclusive results. In order to establish CAL as a viable stem cell therapeutic approach, it will be necessary to demonstrate its oncologic safety in basic and clinical studies. Doing so could transform the paradigm of clinical strategy and practice for the treatment of a wide variety of diseases.

• Archives of Plastic Surgery
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• 제39권5호
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• pp.534-539
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• 2012

Background Autologous fat grafting evolved over the twentieth century to become a quick, safe, and reliable method for restoring volume. However, autologous fat grafts have some problems including uncertain viability of the grafted fat and a low rate of graft survival. To overcome the problems associated with autologous fat grafts, we used uncultured adipose tissue-derived stromal cell (stromal vascular fraction, SVF) assisted autologous fat grafting. Thus, the purpose of this study was to evaluate the effect of SVF in a clinical trial. Methods SVF cells were freshly isolated from half of the aspirated fat and were used in combination with the other half of the aspirated fat during the procedure. Between March 2007 and February 2008, a total of 9 SVF-assisted fat grafts were performed in 9 patients. The patients were followed for 12 weeks after treatment. Data collected at each follow-up visit included clinical examination of the graft site(s), photographs for historical comparison, and information from a patient questionnaire that measured the outcomes from the patient perspective. The photographs were evaluated by medical professionals. Results Scores of the left facial area grafted with adipose tissue mixed with SVF cells were significantly higher compared with those of the right facial area grafted with adipose tissue without SVF cells. There was no significant adverse effect. Conclusions The subjective patient satisfaction survey and surgeon survey showed that SVF-assisted fat grafting was a surgical procedure with superior results.

• Journal of Korean Neurosurgical Society
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• 제40권4호
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• pp.267-272
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• 2006

Objective : There have been recent reports that mesenchymal stromal cells that are harvested from adipose tissue are able to differentiate into neurons. In the present study, we administered adipose tissue derived stem cells in rats with cerebral infarction in order to determine whether those stem cells could enhance the recovery of motor function. Methods : Cerebral infarction was induced by intraluminal occlusion of middle cerebral artery in rats. The adipose tissue-derived mesenchymal stem cells were harvested from inguinal fat pad and proliferated for 2 weeks in DMEM media. Approximately $1{\times}10^6$ cells were injected intravenously or into subdural space of the peri-lesional area. The rotor rod test was performed at preoperative state[before MCA occlusion], and 1, 2, 3, 4, 6, 8 and 10 weeks after the cell therapy. Results : The motor functions that were assessed by rotor rod test at 1 week of the cell therapy were nearly zero among the experimental groups. However, there was apparent motor function recovery after 2 weeks and 4 weeks of cell injection in intravenously treated rats and peri-lesionaly treated rats, respectively, while there was no significant improvement till 8 weeks in vehicle treated rats. Conclusion : These results demonstrate that the adipose derived stem cell treatment improves motor function recovery in rats with cerebral infarction.

• The Korean Journal of Physiology and Pharmacology
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• 제18권4호
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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.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권2호
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• pp.97-106
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• 2010

Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

• The Journal of Korean Physical Therapy
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• 제21권3호
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• pp.87-93
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• 2009

Purpose: TThis study examined the effect of repetitive magnetic stimulation (RMS) on the viability and proliferative response of human adipose tissue-derived stromal cells (hATSCs) in vitro. Methods: The hATSCs were cultured primarily from human adipose tissue harvested by liposuction and incubated in a $37^{\circ}C$ plastic chamber. The cells were exposed to a repetitive magnetic field using a customized magnetic stimulator (Biocon-5000, Mcube Technology). The RMS parameters were set as follows: repetition rate=10Hz, 25Hz (stimulus intensity 100%= 0.1 Tesla, at 4cm from the coil), stimulated time= 1, 5, and 20 minutes. Twenty four hours after one application of RMS, the hATSCs were compared with the sham stimulation, which were kept under the same conditions without the application of RMS. The cells were observed by optical microscopy to determine the morphology and assessed by trypan blue staining for cell proliferation. The apoptosis and viability of the hATSCs were also analyzed by fluorescence-activated cell sorting (FACS) analysis of Annexin V and MTT assay. Results: After RMS, the morphology of the hATSCs was not changed and the apoptosis of hATSCs were not increased compared to the sham stimulation. The viability of the cells was similar to the cells given the sham stimulation. Interestingly, the level of hATSC proliferation was significantly higher in all RMS groups. Conclusion: The application of RMS may not cause a change in morphology and viability of hATSCs but can increase the level of cell proliferation in vitro. RMS might be useful as an adjuvant tool in combination with stem cell therapy without adverse effects.