Objective To investigate the possibility of differentiation of theisolated and cultured adipose-derived adult stem cells into chondrocytes, which is induced by the recombinant human bone morphogenetic protein 2 (rhBMP-2). Methods The rabbit adipose tissue was minced and digested by collagenase Type Ⅰ. The adposederived adult stem cells were obtained and then they were cultured inthe micropellet condition respectively in the rhBMP-2 group, the rhTGF-β1 group, the combination group, and the control group for 14 days. The differentiation of the adiposederived stem cells into chondrocytes was identifiedby the histological methods including HE, Alcian blue, Von kossa, and immunohistochemical stainings. Results After the continuous induction by rhBMP-2 and continuous culture for 14 days, the HE staining revealed a formation of the cartilage lacuna; Alcian blue indicated that proteoglycan existed in the extracellular matrix; the immunohistochemical staining indicated that collagen Ⅱ was in the cellular matrix; and Von kossa indicated that the adipose-derived stem cells couldnot differentiate into the osteoblasts by an induction of rhBMP-2. Conclusion In the micropellet condition, the adipose-derived adult stemcells can differentiate into the chondrocytes, which is initially induced by rhBMP-2. This differentiation can provide a foundation for the repair of the cartilage injury.
Objective To review research progress of adipose tissuederived stromal cells (ADSCs).Methods The recent articles on ADSCs were extensively reviewed, and the culture and differentiation ability of ADSCs were investigated.Results A population of stem cells could be isolated from adult adipose tissue, they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling. The majority of the isolated cells were mesenchymal origin, with a few pericytes,endothelial cells and smooth muscle cells. ADSCs could be induced to differentiate intomultiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic and adipogenic cells, they could also differentiate into nerve cells.Conclusion ADSCs can substitute mesenchymal stem cells and become an alternative stem cells source for tissue engineering.
To isolate and culture adi pose-derived stem cells (ADSCs), and to study the effects of the conditioned medium of ADSCs (ADSC-CM) treated with insul in on HaCaT cells. Methods ADSCs were isolated from adipose tissue donated by the patient receiving abdominal surgery and were cultured. The concentration of ADSCs at passage 3 was adjusted to 5 × 104 cells/mL. The cells were divided into 2 groups: group A in which the cells were incubated in 1 × 10-7 mol/ Linsul in for 3 days, and group B in which the cells were not treated with insul in. ADSC-CM in each group was collected 3 days after culture, then levels of VEGF and hepatocyte growth factor (HGF). HaCaT cells were cultured and the cells at passage 4 were divided into 4 groups: group A1, 0.5 mL 2% FBS and 0.5 mL ADSC-CM from group A; group B1, 0.5 mL 2% FBS and 0.5 mL ADSC-CM from group B; group C1, 1 mL 2% FBS of 1 × 10-7 mol/ L insul in; group D1, 1 mL 2%FBS. Prol iferation of HaCaT cells was detected by MTT method 3 days after culture, apoptosis rate of HaCaT cells was measured by Annexin V-FITC double staining 12 hours after culture, and the migration abil ity was measured by in vitro wound-heal ing assay 0, 12, 24, 36 and 48 hours after culture. Results The level of VEGF in groups A and B was (643.28 ± 63.57) and (286.52 ± 46.68) pg/mL, respectively, and the level of HGF in groups A and B was (929.95 ± 67.52) and (576.61 ± 84.29) pg/mL, respectively, suggesting differences were significant between two groups (Plt; 0.05). Cell prol iferation detection showed the absorbance value of HaCaT cells in group A1, B1, C1 and D1 was 0.881 ± 0.039, 0.804 ± 0.041, 0.663 ± 0.027 and 0.652 ± 0.042, respectively, suggesting there was significant difference between groups A1 and B1 and groups C1 and D1 (P lt; 0.01), group A1 was significantly higher than group B1 (P lt; 0.05). The apoptosis rate of HaCaT cells in groups A1, B1, C1 and D1 was 5.23% ± 1.98%, 8.82% ± 2.59%, 31.70% ± 8.85% and 29.60% ± 8.41%, respectively, indicating there was significant difference between groups A1 and B1 and groups C1 and D1 (P lt; 0.05), group B1 was significantly higher than group A1 (P lt; 0.05). The migration distance of HaCaT cells in groups A1, B1,C1 and D1 at 36 hours was (0.184 6 ± 0.019 2), (0.159 8 ± 0.029 4), (0.059 2 ± 0.017 6) and (0.058 2 ± 0.012 3) mm, respectively, whereas at 48 hours, it was (0.231 8 ± 0.174 0), (0.205 1 ± 0.012 1), (0.079 2 ± 0.008 1) and (0.078 4 ± 0.011 7) mm, respectively, suggesting there were significant differences between groups A1 and B1 and groups C1 and D1 at 36 and 48 hours (P lt; 0.01), group A1 was significantly higher than group B1 (P lt; 0.05) at 36 and 48 hours, no significant difference was evident at other time points(P gt; 0.05). Conclusion ADSCs treated with insul in can significantly promote the prol iferation and the migration of HaCaT cells and inhibit their apoptosis.
ObjectiveTo review the research progress of constructing injectable tissue engineered adipose tissue by adipose-derived stem cells (ADSCs). MethodsRecent literature about ADSCs composite three-dimensional scaffold to construct injectable tissue engineered adipose tissue is summarized, mainly on the characteristics of ADSCs, innovation of injectable scaffold, and methods to promote blood supply. ResultsADSCs have a sufficient amount and powerful ability such as secretion, excellent compatibility with injectable scaffold, plus with methods of promoting blood supply, which can build forms of injectable tissue engineered adipose tissue. ConclusionIn despite of many problems to be dealt with, ADSCs constructing injectable tissue engineered adipose tissue may provide a promising source for soft-tissue defect repair and plastic surgery.
Objective To investigate the possibility of theadipose tissue-derived stromal cells(ADSCs) to differentiate into the neuron-like cells and to explore a new cell source for the transplantation related to the central nervous system. Methods Adipose was digested by collagenase, cultured in the fetal bovine serum containing a medium. Trypse was used to digest the cells and the cell passage was performed. The 3rd to the 9th passage ADSCs were used to make an induction. Isobutylmethylxanthine, indomethacin, insulin, and dexamethasone were used to induce the ADSCs to differentiate into the neuron-like cells and adipocytes. Sudan black B and immunocytochemistry were used to identify the cells. Results A population of the ADSCs could be isolated from the adult human adipose tissue, they were processed to obtain a fibroblast-like population of the cells and could be maintained in vitro for an extendedperiod with the stable population doubling, and they were expanded as the undifferentiated cells in culture for more than 20 passages, which indicated their proliferative capacity. They expressed vimentin and nestin, and characteristics of the neuron precursor stem cells at an early stage of differentiation. And the majority of the ADSCs also expressed the neuron-specific enolase and βⅢ-tubulin, characteristics of the neurons. Isobutyl-methyxanthine, indomethacin, insulin, and dexamethasone induced 40%-50% of ADSCs to differentiate into adipocytes and 0.1%0.2% of ADSCs into neuron-like cells. The neuron-like cells had a complicated morphology of the neurons, and they exhibited a neuron phenotype, expressed nestin, vimentin, neuron-specific enolase and βⅢ-tubulin, but some neuron-like cells also expressed thesmooth muscle actin (SMA), and the characteristics of the smooth muscle cells; however, the neurons from the central nervous system were never reported to express this kind of protein. Therefore, the neuron-like cells from the ADSCs could be regarded as functional neurons. Conclusion Ourresults support the hypothesis that the adult adipose tissue contains the stem cells capable of differentiating into the neuron-like cells, and they can overcome their mesenchymal commitment, which represents an alternative autologous stemcell source for transplantation related to the central nervous system.
Objective To review the mechanism of improved revascularization of free fat grafting with adipose-derived stem cells (ADSCs). Methods The literature related to the basic researches of ADSCs in free fat grafting and angiogenesis was reviewed. Results Angiogenesis is a sequence process in time and space which is regulated by various factors. ADSCs possess the capability of secreting many angiogenic growth factors and differentiating into various lineages.Conclusion ADSCs affect every process of angiogenesis with clear improved angiogenic effects, however, the mechanisms of angiogenic effects need the further researches.
Objective To review the study on adi pose derived stem cells (ADSCs) in the therapy of urological diseases. Methods The recent l iterature concerning ADSCs in bladder repair, urethral reconstruction, incontinence treatment, and erectile dysfunction treatment was reviewed. Results The appl ication of tissue engineering using ADSCs has made significant achievements in the treatment of urological diseases and in animal studies, and has been initially used in cl inicaland has achieved a good therapeutic effect. Conclusion Tissue engineering using ADSCs has good prospects in the study on urological diseases, and is expected to widely used in the treatment of urological diseases.
Objective To investigate the differentiation of theadipose-derived adult stem cell (ADASC) induced by the recombinant adenovirus’s containing fibers derived from B-group serotype 35 (rAd5/F35)mediated human bone morphogenetic protein 7 (hBMP-7) gene and to explore a new cell sourcefor the bone tissue engineering. Methods The hBMP-7 gene wasamplified with the pcDNA1.1/AMP-hBMP-7 plasmid as a formwork. After the purification, the gene fragment was cloned into the pDC316 carrier for the recombination of the plasmid of pDC316-hBMP-7. The 293 cells were cotransfected by the skeleton plasmid of pBHG-fiber5/35 and the shuttle plasmid of pDC316-hBMP-7, and the recombinant plasmid of Ad5/F35-hBMP-7 was obtained; the recombinant plasmid of Ad5/F35enhancd green fluorescent protein(EGFP) was obtained by the similar method. The rat ADASCs were cultured and transfected by the Ad5/F35-hBMP-7plasmid and the Ad5/F35-EGFP plasmid, respectively; the remaining untransfected ADASC were used as the controls. The morphology and the growth pattern of the transfected cells were evaluated. The transcription and the expression of the transfected genes and the steogenic phenotypes such as calcium nodules and osteocalcin were evaluated by ELISA. Results The identification of PCR and enzyme cutting showed that the construction of the recombinant Ad5/F35-hBMP-7 plasmid could be confirmed. The transfection rate of the ADASC by the Ad5/F35-EGFP plasmid was determined to be greater than 90%. The hBMP-7 gene in thetransfected ADASC could express the corresponding protein, and the formation ofthe calcium nodules could be found in the induced group. The electron microscopy showed that there was a calcium element in the cytoplasm, the alkaline phosphatase result was positive, and the expression of osteocalcin was increased. Conclusion The rAd5/F35-hBMP-7 gene can promote the differentiation of the adiposederived adult stem cells to the osteoblasts in the bone tissue engineering.
Objective To introduce types and differentiation potentials of stem cells from adipose tissue, and its applications on regenerative medicine and advantages. Methods The literature of original experimental study and clinical research about bone marrow mesenchymal stem cells (BMSCs), adipose-derived stem cells (ADSCs), and dedifferentiated fat (DFAT) cells was extensively reviewed and analyzed. Results ADSCs can be isolated from stromal vascular fraction. As ADSCs have multi-lineage potentials, such as adipogenesis, osteogenesis, chondrogenesis, angiogenesis, myogenesis, and neurogenesis, they have already been successfully used in regenerative medicine areas. Dramatically, mature fat cells can be dedifferentiated and changed into fibroblast-like cells, named DFAT cells, via ceiling culture method. DFAT cells also had the same multi-lineage potentials as ADSCs, differentiating into adipocytes, osteocytes, chondrocytes, endothelial cells, muscle cells, and nerve cells. Compared with BMSCs which are commonly used as adult stem cells, ADSCs and DFAT cells have extensive sources and can be easily acquired. While compared with ADSCs, DFAT cells have good homogeneity and b proliferation capacity. Conclusion As a potential source of stem cells, adipose tissue will provide a new promising for regenerative medicine.
Objective To review the biochemical characteristics, appl ication progress, and prospects of the adiposederived stem cells (ADSCs). Methods The recent original experimental and cl inical l iterature about ADSCs was extensively reviewed and analyzed. Results ADSCs can be readily harvested in large numbers from adipose tissue with properties of stable prol iferation and potential differentiation in vitro. Significant progress of ADSCs is made in the animal experimentand the cl inical appl ication. It has been widely used in the cl inical treatment of cardiovascular disease, metabol ic disease, encephalopathy, and tissue engineering repair. Conclusion ADSCs have gradually replaced bone marrow mesenchymal stem cells and become the focused hot spot of regenerative medicine and stem cells.