Adipose tissue-derived stem cells as a therapeutic tool for cardiovascular disease

Adipose tissue-deried stem cells( ADSCs) are adult stem cells that can be easily harvested from subcutaneous adipose tissue. Many studies have demonstrated that ADSCs differentiate into vascular endothelial cells(VECs), vascular smooth muscle cells(VSMCs), and cardiomyocytes in vitro and in vivo. However, ADSCs may fuse with tissue-resident cells and obtain the corresponding characteristics of those cells. If fusion occurs, ADSCs may express markers of VECs, VSMCs, and cardiomyocytes without direct differentiation into these cell types. ADSCs also produce a variety of paracrine factors such as vascular endothelial growth factor, hepatocyte growth factor, and insulin-like growth factor-1 that have proangiogenic and/or antiapoptotic activities. Thus, ADSCs have the potential to regenerate the cardiovascular system via direct differentiation into VECs, VSMCs, and cardiomyocytes, fusion with tissueresident cells, and the production of paracrine factors. Numerous animal studies have demonstrated the efficacy of ADSC implantation in the treatment of acute myocardial infarction(AMI), ischemic cardiomyopathy(ICM), dilated cardiomyopathy, hindlimb ischemia, and stroke. Clinical studies regarding the use of autologous ADSCs for treating patients with AMI and ICM have recently been initiated. ADSC implantation has been reported as safe and effective so far. Therefore, ADSCs appear to be useful for the treatment of cardiovascular disease. However, the tumorigenic potential of ADSCs requires careful evaluation before their safe clinical application.

High-Fat Diets-Induced Metabolic Alterations Alter the Differentiation Potential of Adipose Tissue-Derived Stem Cells

Background: Adipose tissue-derived stem cells (ASC) possess the ability to differentiate into adipocytes or endothelial cells to help in the adipogenesis, vasculogenesis and vascular repair. This study aims at determining the impact of high-fat diets (HFD)-induced type 2 diabetes (T2D) on the differentiation potential of ASC. Results: C57BL/6J male mice were fed a vegetal (VD) or an animal (AD) HFD. Isolation of ACS from mice showing different levels of metabolic alterations reveals that advanced T2D did not affect the number of cells per gram of tissue. Rather, a higher proportion of inflammatory CD36+ cells was identified in HFD fed mice. Despite a marked decreased expression of adipogenic genes (aP2, C/EBPα and PPARγ2), ASC from HFD groups had a higher adipogenic potential and a lower endothelial differentiation potential in vitro compared to control. ASC from the VD group had enhanced cyclin B1 expression and had more adipogenic potential compared to AD group. Conclusion: Our results demonstrate that the metabolic modifications, linked to the nature of fatty acids in diets, modulate the differentiation potential of ASC with increased adipogenesis to the detriment of the endothelial pathway. Results highlight the importance of evaluating the ASC differentiation behavior in a context of autologous cell-based therapy for the repair of vascular tissues in diabetic patients.

Optimization of adipose tissue-derived mesenchymal stromal cells transplantation for bone marrow repopulation following irradiation

BACKGROUND Bone marrow(BM)suppression is one of the most common side effects of radiotherapy and the primary cause of death following exposure to irradiation.Despite concerted efforts,there is no definitive treatment method available.Recent studies have reported using mesenchymal stromal cells(MSCs),but their therapeutic effects are contested.AIM We administered and examined the effects of various amounts of adipose-derived MSCs(ADSCs)in mice with radiation-induced BM suppression.METHODS Mice were divided into three groups:Normal control group,irradiated(RT)group,and stem cell-treated group following whole-body irradiation(WBI).Mouse ADSCs(mADSCs)were transplanted into the peritoneal cavity either once or three times at 5×10^(5) cells/200μL.The white blood cell count and the levels of,plasma cytokines,BM mRNA,and BM surface markers were compared between the three groups.Human BM-derived CD34+hematopoietic progenitor cells were co-cultured with human ADSCs(hADSCs)or incubated in the presence of hADSCs conditioned media to investigate the effect on human cells in vitro.RESULTS The survival rate of mice that received one transplant of mADSCs was higher than that of mice that received three transplants.Multiple transplantations of ADSCs delayed the repopulation of BM hematopoietic stem cells.Anti-inflammatory effects and M2 polarization by intraperitoneal ADSCs might suppress erythropoiesis and induce myelopoiesis in sub-lethally RT mice.CONCLUSION The results suggested that an optimal amount of MSCs could improve survival rates post-WBI.

Outcomes of combined mitochondria and mesenchymal stem cellsderived exosome therapy in rat acute respiratory distress syndrome and sepsis

BACKGROUND The treatment of acute respiratory distress syndrome(ARDS)complicated by sepsis syndrome(SS)remains challenging.AIM To investigate whether combined adipose-derived mesenchymal-stem-cells(ADMSCs)-derived exosome(EXAD)and exogenous mitochondria(mitoEx)protect the lung from ARDS complicated by SS.METHODS In vitro study,including L2 cells treated with lipopolysaccharide(LPS)and in vivo study including male-adult-SD rats categorized into groups 1(sham-operated-control),2(ARDS-SS),3(ARDS-SS+EXAD),4(ARDS-SS+mitoEx),and 5(ARDS-SS+EXAD+mitoEx),were included in the present study.RESULTS In vitro study showed an abundance of mitoEx found in recipient-L2 cells,resulting in significantly higher mitochondrial-cytochrome-C,adenosine triphosphate and relative mitochondrial DNA levels(P<0.001).The protein levels of inflammation[interleukin(IL)-1β/tumor necrosis factor(TNF)-α/nuclear factor-κB/toll-like receptor(TLR)-4/matrix-metalloproteinase(MMP)-9/oxidative-stress(NOX-1/NOX-2)/apoptosis(cleaved-caspase3/cleaved-poly(ADP-ribose)polymerase)]were significantly attenuated in lipopolysaccharide(LPS)-treated L2 cells with EXAD treatment than without EXAD treatment,whereas the protein expressions of cellular junctions[occluding/β-catenin/zonula occludens(ZO)-1/E-cadherin]exhibited an opposite pattern of inflam-mation(all P<0.001).Animals were euthanized by 72 h post-48 h-ARDS induction,and lung tissues were harvested.By 72 h,flow cytometric analysis of bronchoalveolar lavage fluid demonstrated that the levels of inflam-matory cells(Ly6G+/CD14+/CD68+/CD11b/c+/myeloperoxidase+)and albumin were lowest in group 1,highest in group 2,and significantly higher in groups 3 and 4 than in group 5(all P<0.0001),whereas arterial oxygen-saturation(SaO2%)displayed an opposite pattern of albumin among the groups.Histopathological findings of lung injury/fibrosis area and inflammatory/DNA-damaged markers(CD68+/γ-H2AX)displayed an identical pattern of SaO2%among the groups(all P<0.0001).The protein expressions of inflammatory(TLR-4/MMP-9/IL-1β/TNF-α)/oxidative stress(NOX-1/NOX-2/p22phox/oxidized protein)/mitochondrial-damaged(cytosolic-cytochrome-C/dynamin-related protein 1)/autophagic(beclin-1/Atg-5/ratio of LC3B-II/LC3B-I)biomarkers exhibited a similar manner,whereas antioxidants[nuclear respiratory factor(Nrf)-1/Nrf-2]/cellular junctions(ZO-1/E-cadherin)/mitochondrial electron transport chain(complex I-V)exhibited an opposite manner of albumin among the groups(all P<0.0001).CONCLUSION Combined EXAD-mitoEx therapy was better than merely one for protecting the lung against ARDS-SS induced injury.

Recent advances in andrology-related stem cell research

Stem cells hold great promise for regenerative medicine because of their ability to self-renew and to differentiate into various cell types. Although embryonic stem cells （BSC） have greater differentiation potential than adult stem cells, the former is lagging in reaching clinical applications because of ethical concerns and governmental restrictions. Bone marrow stem cells （BMSC） are the best-studied adult stem cells （ASC） and have the potential to treat a wide variety of diseases, including erectile dysfunction （ED） and male infertility. More recently discovered adipose tissuederived stem cells （ADSC） are virtually identical to bone marrow stem cells in differentiation and therapeutic potential, but are easier and safer to obtain, can be harvested in larger quantities, and have the associated benefit of reducing obesity. Therefore, ADSC appear to be a better choice for future clinical applications. We have previously shown that ESC could restore the erectile function of neurogenic ED in rats, and we now have evidence that ADSC could do so as well. We are also investigating whether ADSC can differentiate into Leydig, Sertoli and male germ cells. The eventual goal is to use ADSC to treat male infertility and testosterone deficiency. （Asian JAndrol 2008 Mar; 10： 171-175）

Over-expression of VEGF165 in the adipose tissue-derived stem cells via the lentiviral vector

Background Many researchers studied the possibility of using stem cells as gene therapeutic vector. But few related reports on the adipose tissue-derived stem cells （ADSCs） are available. Therefore we intended to construct a lentiviral VEGF165 expression vector and then infect the ADSCs to produce therapeutic seed cells.Methods EHS1001-68950485313912 clone was mutated by PCR method to produce consensus fragment of VEGF165 transcript （NM_001025368）. Lentivirus was enveloped with pGC-FU, pHelper 1.0 and pHelper 2.0 plasmids in 293T cells.And then the ADSCs （multiplicity of infection=20） were transfected with the vectors after titer determination. Stable expression of VEGF165 in ADSCs was confirmed by immunofluorescence staining, enzyme-linked immunosorbent assay （ELISA） and Western blotting analysis.Results DNA sequencing and 293T transfection verified VEGF165 was linked to the GFP fused vector. The virus titer is up to 2x10a determined by quantitative PCR. VEGF165 transduced cells could show green fluorescence confirmed by immunofluorescence staining （almost 95%）. ELISA analyses could detect out the density of VEGF was 850.86-1202.13pg/ml （mean （923.00±31.22） pg/ml） in the supernatant of VEGF16s-transduced cells but not detected in the GFP-transduced cells （P 〈0.001） and the Western blotting analyses also confirmed VEGF165 expression in VEGF165-transduced cells.Conclusions The VEGF165 over-expression ADSCs were obtained and may be used as a cell therapeutic tool and may be applied for vascular regeneration, especially in the treatment of erectile dysfunction.

Stem cells： novel players in the treatment of erectile dysfunction

Stem cells are defined by their capacity for both self-renewal and directed differentiation; thus, they represent great promise for regenerative medicine. Historically, stem cells have been categorized as either embryonic stem cells （ESCs） or adult stem cells （ASCs）. It was previously believed that only ESCs hold the ability to differentiate into any cell type, whereas ASCs have the capacity to give rise only to cells of a given germ layer. More recently, however, numerous studies demonstrated the ability of ASCs to differentiate into cell types beyond their tissue origin. The aim of this review was to summarize contemporary evidence regarding stem cell availability, differentiation, and more specifically, the potential of these cells in the diagnosis and treatment of erectile dysfunction （ED） in both animal models and human research. We performed a search on PubMed for articles related to definition, iocalisation and circulation of stem cells as well as the application of stem cells in both diagnosis and treatment of ED. Strong evidence supports the concept that stem cell therapy is potentially the next therapeutic approach for ED. To date, a large spectrum of stem cells, including bone marrow mesenchymal stem cells, adipose tissue-derived stem cells and muscle-derived stem cells, have been investigated for neural, vascular, endothelial or smooth muscle regeneration in animal models for ED. In addition, several subtypes of ASCs are localized in the penis, and circulating endogenous stem cells can be employed to predict the outcome of ED and ED-related cardiovascular diseases.

Rho-associated coiled kinase inhibitor Y-27632 promotes neuronal-like differentiation of adult human adipose tissue-derived stem cells

Background Y-27632 is a specific inhibitor of Rho-associated coiled kinase （ROCK） and has been shown to promote the survival and induce the differentiation of a variety of cells types. However, the effects of Y-27632 on adult human adipose tissue-derived stem cells （ADSCs） are unclear. This study aimed to investigate the effects of Y-27632 on the neuronal-like differentiation of ADSCs. Methods ADSCs were isolated from women undergoing plastic surgery and cultured. ADSCs were treated with different doses of Y-27632 and observed morphological changes under microscope. The expression of nestin, neuron specific enolase （NSE） and microtubule-associated protein-2 （MAP-2） in ADSCs treated with Y-27632 was detected by immunocytochemistry and Western blotting analysis. Results Y-27632 had the potency to induce neuronal-like differentiation in ADSCs in a dose-dependent manner. Moreover, the differentiation induced by Y-27632 was recovered upon drug withdraw. ADSCs treated with Y-27632 expressed neuronal markers such as NSE, MAP-2 and nestin while untreated ADSCs did not express these markers. Conclusion Selective ROCK inhibitor Y-27632 could potentiate the neuronal-like differentiation of ADSCs, suggesting that Y-27632 could be utilized to induce the differentiation of ADSCs to neurons and facilitate the clinical application of ADSCs in tissue engineering.

Chemically induced hypoxia by dimethyloxalylglycine(DMOG)-loaded nanoporous silica nanoparticles supports endothelial tube formation by sustained VEGF release from adipose tissue-derived stem cells

Inadequate vascularization leading to insufficient oxygen and nutrient supply in deeper layers of bioartificial tissues remains a limitation in current tissue engineering approaches to which prevascularization offers a promising solution.Hypoxia triggering pre-vascularization by enhanced vascular endothelial growth factor(VEGF)expression can be induced chemically by dimethyloxalylglycine(DMOG).Nanoporous silica nanoparticles(NPSNPs,or mesoporous silica nanoparticles,MSNs)enable sustained delivery of molecules and potentially release DMOG allowing a durable capillarization of a construct.Here we evaluated the effects of soluble DMOG and DMOG-loaded NPSNPs on VEGF secretion of adipose tissue-derived stem cells(ASC)and on tube formation by human umbilical vein endothelial cells(HUVEC)-ASC co-cultures.Repeated doses of 100 mM and 500 mM soluble DMOG on ASC resulted in 3-to 7-fold increased VEGF levels on day 9(P<0.0001).Same doses of DMOG-NPSNPs enhanced VEGF secretion 7.7-fold(P<0.0001)which could be maintained until day 12 with 500 mM DMOG-NPSNPs.In fibrin-based tube formation assays,100 mM DMOG-NPSNPs had inhibitory effects whereas 50 mM significantly increased tube length,area and number of junctions transiently for 4 days.Thus,DMOG-NPSNPs supported endothelial tube formation by upregulated VEGF secretion from ASC and thus display a promising tool for prevascularization of tissue-engineered constructs.Further studies will evaluate their effect in hydrogels under perfusion.

Unmodified autologous stem cells at point of care for chronic myocardial infarction

BACKGROUND Numerous studies investigated cell-based therapies for myocardial infarction(MI).The conflicting results of these studies have established the need for developing innovative approaches for applying cell-based therapy for MI.Experimental studies on animal models demonstrated the potential of fresh,uncultured,unmodified,autologous adipose-derived regenerative cells(UAADRCs)for treating acute MI.In contrast,studies on the treatment of chronic MI(CMI;>4 wk post-MI)with UA-ADRCs have not been published so far.Among several methods for delivering cells to the myocardium,retrograde delivery into a temporarily blocked coronary vein has recently been demonstrated as an effective option.AIM To test the hypothesis that in experimentally-induced chronic myocardial infarction(CMI;>4 wk post-MI)in pigs,retrograde delivery of fresh,uncultured,unmodified,autologous adipose-derived regenerative cells(UA-ADRCs)into a temporarily blocked coronary vein improves cardiac function and structure.METHODS The left anterior descending(LAD)coronary artery of pigs was blocked for 180 min at time point T0.Then,either 18×106 UA-ADRCs prepared at“point of care”or saline as control were retrogradely delivered via an over-the-wire balloon catheter placed in the temporarily blocked LAD vein 4 wk after T0(T1).Effects of cells or saline were assessed by cardiac magnetic resonance(CMR)imaging,late gadolinium enhancement CMR imaging,and post mortem histologic analysis 10 wk after T0(T2).RESULTS Unlike the delivery of saline,delivery of UA-ADRCs demonstrated statistically significant improvements in cardiac function and structure at T2 compared to T1(all values given as mean±SE):Increased mean LVEF(UA-ADRCs group:34.3%±2.9%at T1 vs 40.4±2.6%at T2,P=0.037;saline group:37.8%±2.6%at T1 vs 36.2%±2.4%at T2,P>0.999),increased mean cardiac output(UA-ADRCs group:2.7±0.2 L/min at T1 vs 3.8±0.2 L/min at T2,P=0.002;saline group:3.4±0.3 L/min at T1 vs 3.6±0.3 L/min at T2,P=0.798),increased mean mass of the left ventricle(UA-ADRCs group:55.3±5.0 g at T1 vs 71.3±4.5 g at T2,P<0.001;saline group:63.2±3.4 g at T1 vs 68.4±4.0 g at T2,P=0.321)and reduced mean relative amount of scar volume of the left ventricular wall(UA-ADRCs group:20.9%±2.3%at T1 vs 16.6%±1.2%at T2,P=0.042;saline group:17.6%±1.4%at T1 vs 22.7%±1.8%at T2,P=0.022).CONCLUSION Retrograde cell delivery of UA-ADRCs in a porcine model for the study of CMI significantly improved myocardial function,increased myocardial mass and reduced the formation of scar tissue.

Stem cell therapy for erectile dysfunction

Erectile dysfunction(ED)is an important health problem that has commonly been clinically treated using phosphodiesterase type 5 inhibitors(PDE5Is).However,PDE5Is are less effective when the structure of the cavernous body has been severely injured,and thus regeneration is required.Stem cell therapy has been investigated as a possible means for regenerating the injured cavernous body.Stem cells are classified into embryonic stem cells and adult stem cells(ASCs),and the intracavernous injection of ASCs has been explored as a therapy in animal ED models.Bone marrowderived mesenchymal stem cells and adipose tissuederived stem cells are major sources of ASCs used for the treatment of ED,and accumulated evidence now suggests that ASCs are useful in the restoration of erectile function and the regeneration of the cavernous body.However,the mechanisms by which ASCs recover erectile function remain controversial.Some studies indicated that ASCs were differentiated into the vascular endothelial cells,vascular smooth muscle cells,and nerve cells that originally resided in the cavernous body,whereas other studies have suggested that ASCs improved erectile function via the secretion of anti-apoptotic and/or proangiogenic cytokines ratherthan differentiation into other cell types.In this paper,we reviewed the characteristics of stem cells used for the treatment of ED,and the possible mechanisms by which these cells exert their effects.We also discussed the problems to be solved before implementation in the clinical setting.

Transplantation of neural progenitor cells differentiated from adipose tissue-derived stem cells for treatment of sciatic nerve injury

Objectives: Currently, the clinical repair of sciatic nerve injury remains difficult.Previous studies have confirmed that transplantation of adipose tissue-derived stem cells promotes nerve regeneration and restoration at peripheral nerve injury sites. Methods: In this study, adipose tissue-derived stem cells were induced to differentiate into neural progenitor cells, transfected with a green fluorescent protein-containing lentivirus, and then transplanted into the lesions of rats with sciatic nerve compression injury. Results: Fluorescence microscopy revealed that the transplanted cells survived,migrated, and differentiated in rats. At two weeks post-operation, a large number of transplanted cells had migrated to the injured lesions; at six weeks post-operation, transplanted cells were visible around the injured nerve and several cells were observed to express a Schwann cell marker. Sciatic function index and electrophysiological outcomes of the transplantation group were better than those of the control group. Cell transplantation promoted the recovery of motor nerve conduction velocity and compound muscle action potential amplitude, and reduced gastrocnemius muscle atrophy.Conclusions: Our experimental findings indicate that neural progenitor cells,differentiated from adipose tissue-derived stem cells, are potential seed stem cells that can be transplanted into lesions to treat sciatic nerve injury. This provides a theoretical basis for their use in clinical applications.

Metformin promotes angiogenesis by enhancing VEGFa secretion by adipose-derived stem cells via the autophagy pathway

Human adipose tissue-derived stem cell(ADSC)derivatives are cell-free,with low immunogenicity and no potential tumourigenicity,making them ideal for aiding wound healing.However,variable quality has impeded their clinical application.Metformin(MET)is a 5′adenosine monophosphate-activated protein kinase activator associated with autophagic activation.In this study,we assessed the potential applicability and underlying mechanisms of MET-treated ADSC derivatives in enhancing angiogenesis.We employed various scientific techniques to evaluate the influence of MET on ADSC,assess angiogenesis and autophagy in MET-treated ADSC in vitro,and examine whether MET-treated ADSC increase angiogenesis.We found that low MET concentrations exerted no appreciable effect on ADSC proliferation.However,MET was observed to enhance the angiogenic capacity and autophagy of ADSC.MET-induced autophagy was associated with increased vascular endothelial growth factor A production and release,which contributed to promoting the therapeutic efficacy of ADSC.In vivo experiments confirmed that in contrast to untreated ADSC,MET-treated ADSC promoted angiogenesis.Our findings thus indicate that the application of MET-treated ADSC would be an effective approach to accelerate wound healing by promoting angiogenesis at wound sites.

Nanotechnology-assisted adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury： In vivo cell tracking, optimized injection dosage, and functional evaluation

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury （CNI） by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage （2 × 105 cells） of ADSCs for intracavernous injection （ICI） and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs （NanoADSCs） were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.

兔脂肪干细胞(ADSCs)与聚羟基乙酸/壳聚糖(PLGA/CS)支架材料生物相容性研究

目的探讨兔脂肪来源于细胞（adipose-derived stemcells，ADSCs）作为种子细胞复合新型聚羟基乙酸（polylaetic-co—glycolicacid，PLGA）／壳聚糖（chitosan，CS）支架材料的生物相容性，为进一步构建组织工程化脂肪组织提供实验依据。方法取雄性新西兰大白兔腹股沟脂肪组织，I型胶原酶消化分离出ADSCs进行培养，贴壁细胞至3～5代，评价其多向分化能力。将干细胞收集重悬后，以l×10^7／mL的密度接种于多孔PLGA／CS支架，形成细胞一支架材料复合物。培养7天后，扫描电子显微镜观察细胞在支架材料上的黏附生长和基质分泌情况，评价细胞与支架材料的生物相容性；Dil荧光标记检测细胞在支架材料上的分布；Hochest33258检测细胞在支架上的生长情况。接种1和7天后，分别对细胞一材料复合物行AnnexinV／PI双染色法检测材料对细胞的毒性作用。用成脂诱导条件培养基诱导分化ADSCs及ADSCs支架材料复合物，7天后，尼罗红荧光染色液检测ADSCs在不同环境下的成脂分化能力。结果原代培养的ADSCs呈成纤维细胞样外观，在相应诱导条件下能够分化为脂肪细胞和骨细胞。细胞接种于PLGA／CS支架材料上第8天分裂增殖达到高峰，扫描电子显微镜下提示ADSCs在支架表面贴附生长良好，并向孔隙内壁充分延伸，细胞周围形成丰富的基质成分。活死双染结合共聚焦显微镜显示材料对细胞活性无影响，尼罗红染色可见成脂诱导后的ADSCs细胞质内有红色脂滴颗粒形成。结论多孔PLGA／CS支架与兔ADSCs具有良好的生物相容性，可作为构建组织工程脂肪组织的支架材料。

白藜芦醇联合施万细胞样细胞对大鼠坐骨神经损伤的修复作用

目的:探讨白藜芦醇联合脂肪源性干细胞(ADSCs)分化的施万细胞样细胞(SCLCs)对大鼠坐骨神经损伤的修复作用。方法:ADSCs原代培养并向SCLCs诱导分化,扫描电镜观察细胞形态;Western Blot方法检测S100钙结合蛋白β(S100β)、p75神经营养素受体(p75NTR)、胶质纤维酸性蛋白(GFAP)表达。大鼠随机分为对照组(Control)、施万细胞样细胞组(SCLCs)、白藜芦醇组(Res)、白藜芦醇+施万细胞样细胞组(Res+SCLCs),坐骨神经损伤模型造模成功后8周通过足迹实验检测各组坐骨神经功能指数(SFI);von Frey丝刺激针测定机械缩足阈值(MWT);称重法测定胫前肌湿重比率(WR);Western Blot、RT-qPCR方法检测损伤处神经营养素-3(NT-3)、神经生长因子(NGF)、类胰岛素生长因子-1(IGF-1)和脑源性神经营养因子(BDNF)表达情况。结果:ADSCs诱导8 d后细胞两极细长,细胞外成分增多;S100β、p75NTR、GFAP蛋白高表达。给予SCLCs、Res及Res+SCLCs治疗后,治疗组SFI、WR明显优于Control组(P<0.05);Res+SCLCs组、SCLCs组大鼠MWT降低(P<0.05)。Western Blot结果显示:Res+SCLCs组大鼠NT-3、IGF-1、NGF、BDNF蛋白表达高于其余各组(P<0.05);SCLCs组大鼠NT-3、NGF、BDNF蛋白表达高于Control组(P<0.05);Res组大鼠NT-3、NGF蛋白表达高于Control组(P<0.05)。RT-qPCR结果显示:Res+SCLCs组大鼠NT-3、IGF-1、NGF、BDNF mRNA高表达;SCLCs组大鼠NT-3、IGF-1、NGF、BDNF mRNA表达高于Control组(P<0.05);Res组大鼠IGF-1、NGF mRNA表达高于Control组(P<0.05)。结论:Res联合ADSCs分化的SCLCs对大鼠坐骨神经损伤具有良好的修复作用。

兔脂肪干细胞MR示踪前的超顺磁性氧化铁最佳标记条件筛选

目的 采用多聚赖氨酸修饰的超顺磁性氧化铁(Polylysine coated superparamagnetic iron oxide,PLL-SPIO)纳米颗粒标记兔脂肪间充质干细胞(Adipose derived stem cell,ADSCs),探讨不同SPIO浓度及孵育时间对ADSCs标记率、存活率及细胞增殖率的影响,筛选SPIO标记ADSCs的最优标记条件。方法 用含不同浓度的SPIO(12、25、50μg/mL)培养基培养第3代兔ADSCs,分别孵育不同时间(6、12、24、48 h),孵育完成后,普鲁士蓝染色计算细胞标记率;细胞毒性实验检测细胞存活率;根据细胞毒性实验结果筛选细胞存活率较大的6组细胞进行细胞增殖能力检测,检测细胞增殖率。结果 普鲁士蓝染色结果显示,各组细胞标记率及细胞内蓝染颗粒数量,随着SPIO浓度增大及孵育时间的延长而增多。细胞毒性实验结果显示,在较低SPIO浓度和较短孵育时间下,细胞存活率不受影响,且各组之间细胞存活率无明显差异,随着SPIO浓度增大及孵育时间延长,细胞存活率呈下降趋势,48 h时细胞存活率下降最明显。细胞增殖实验结果显示,实验组各组细胞增殖率,随SPIO浓度增大及孵育时间延长而降低;且细胞进入快速增长期后,实验组细胞增殖率与阴性对照组相比,均存在明显差异,其中12μg/mL SPIO-6 h组和25μg/mL SPIO-6 h组细胞增殖率与阴性对照组差异较其他各组小。结论 从细胞内SPIO颗粒含量及细胞标记率、存活率及增殖率几方面综合评价,25μg/mL SPIO孵育6 h,是本研究所选出的PLL-SPIO标记ADSCs的最优标记条件。

脂肪干细胞在肥胖症治疗中的研究进展

随着经济的不断发展和生活水平的提高,肥胖症的患病率正逐年显著升高。肥胖症不仅会引起机体能量代谢失衡,还会导致脂肪组织组成细胞的功能紊乱。脂肪干细胞(adipose-derived stemcell,ADSC)是形成脂肪组织的起源细胞,能够改变脂肪组织的重塑和功能,目前被认为可在肥胖症防治中发挥重要作用。为此,本文就ADSC的生物学特征、功能以及其在抗肥胖症中的作用机制进行综述,以期为肥胖症的临床防治研究提供新思路。

脂肪干细胞(ADSCs)在瘢痕防治中的基础研究进展

瘢痕组织是人体创伤过程中的一种自然产物,它不但影响外观,甚至还有可能因为挛缩而引起功能障碍。瘢痕的发生机制尚未完全清楚,目前人体炎症介质过度分泌、细胞因子释放失调、成纤维细胞增殖和降解不平衡以及胶原过度沉积被认为是瘢痕形成的主要因素。瘢痕的治疗方案主要包括抗瘢痕药物的应用、手术切除、皮瓣转移、皮肤软组织扩张、CO2激光等,都具有一定的效果,但亦有一定的局限性。近年来,脂肪干细胞（adi pose derived stem cells,ADSCs）因其特性被大家所重视,现就ADSCs的主要生理特性作一综述。

The CD133/1⁺cell subset from human subcutaneous adult fat retains hemogenic potential

Research has shown that cells from adult fat tissue can effect long-term blood reconstitution. Fat-derived multipotentiality was ascribed to CD34+ perivascular populations from its prominent microvasculature, that represent mostly non-hemogenic, mesenchymal cells, although this tissue contains a CD34+45+ subset committed to a hemogenic fate. Here, in order to analyze cell subsets presenting hemogenic capabilities within fat, CD133/1+ and pericytes, the latter defined by CD140b (PDGFRb, Platelet-Derived Growth Factor Receptor Beta) expression, were immunomagnetically selected from stromal-vascular fractions (SVF). In Vitro Colony Forming Unit (CFU) assays were negative for CD140b+ pericytes and positive for CD133/1+ cells when a prolonged CFU assay was performed, revealing fat as another store of primitive progenitors that retain hemogenic potential.