Therapeutic effects of menstrual blood-derived endometrial stem cells on mouse models of streptozotocin-induced type 1 diabetes

BACKGROUND Type 1 diabetes(T1D),a chronic metabolic and autoimmune disease,seriously endangers human health.In recent years,mesenchymal stem cell(MSC)transplantation has become an effective treatment for diabetes.Menstrual bloodderived endometrial stem cells(MenSC),a novel MSC type derived from the decidual endometrium during menstruation,are expected to become promising seeding cells for diabetes treatment because of their noninvasive collection procedure,high proliferation rate and high immunomodulation capacity.AIM To comprehensively compare the effects of MenSC and umbilical cord-derived MSC(UcMSC)transplantation on T1D treatment,to further explore the potential mechanism of MSC-based therapies in T1D,and to provide support for the clinical application of MSC in diabetes treatment.METHODS A conventional streptozotocin-induced T1D mouse model was established,and the effects of MenSC and UcMSC transplantation on their blood glucose and serum insulin levels were detected.The morphological and functional changes in the pancreas,liver,kidney,and spleen were analyzed by routine histological and immunohistochemical examinations.Changes in the serum cytokine levels in the model mice were assessed by protein arrays.The expression of target proteins related to pancreatic regeneration and apoptosis was examined by western blot.RESULTS MenSC and UcMSC transplantation significantly improved the blood glucose and serum insulin levels in T1D model mice.Immunofluorescence analysis revealed that the numbers of insulin+and CD31+cells in the pancreas were significantly increased in MSC-treated mice compared with control mice.Subsequent western blot analysis also showed that vascular endothelial growth factor(VEGF),Bcl2,Bcl-xL and Proliferating cell nuclear antigen in pancreatic tissue was significantly upregulated in MSC-treated mice compared with control mice.Additionally,protein arrays indicated that MenSC and UcMSC transplantation significantly downregulated the serum levels of interferonγand tumor necrosis factorαand upregulated the serum levels of interleukin-6 and VEGF in the model mice.Additionally,histological and immunohistochemical analyses revealed that MSC transplantation systematically improved the morphologies and functions of the liver,kidney,and spleen in T1D model mice.CONCLUSION MenSC transplantation significantly improves the symptoms in T1D model mice and exerts protective effects on their main organs.Moreover,MSC-mediated angiogenesis,antiapoptotic effects and immunomodulation likely contribute to the above improvements.Thus,MenSC are expected to become promising seeding cells for clinical diabetes treatment due to their advantages mentioned above.

Therapeutic potential of menstrual blood stem cells in treating acute liver failure

BACKGROUND Acute liver failure(ALF)is a significant and complex hepatic insult that may rapidly progress to life-threatening conditions.Recently,menstrual blood stem cells(MenSCs)have been identified as a group of easily accessible mesenchymal stem cells with the advantages of non-invasive acquisition,low immunogenicity,a greater capacity of self-renewal and multi-lineage differentiation,making them promising candidates for stem cell-based therapy to revolutionize the treatment strategies for liver failure.AIM To investigate the therapeutic potential of MenSCs for treating ALF in pigs and to dynamically trace the biodistribution of transplanted cells.METHODS MenSCs were labeled in vitro with PKH26,a lipophilic fluorescent dye.The treatment group received immediate transplantation of PKH26-labelled MenSCs(2.5×106/kg)via the portal vein after D-galactosamine injection,and the control group underwent sham operation.The survival time,liver function,and hepatic pathological changes were compared between the two groups.Three major organs(liver,lungs and spleen)were extracted from animals and imaged directly with the In vivo Imaging System(IVIS)at the predetermined time points.The regions of interest were drawn to quantify the cell uptake in different organs.RESULTS The labelling procedure did not affect the morphology,viability or multipotential differentiation of MenSCs.Biochemical analysis showed that the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL)and prothrombin time(PT)measured at selected time points 24 h after transplantation were significantly decreased in the treatment group(P<0.05).The survival time of ALF animals was prolonged in the treatment group compared with the control group(75.75±5.11 h vs 53.75±2.37 h,log rank,P<0.001).The liver pathological tissue in the MenSC treatment group showed obviously increased numbers of remaining hepatocytes and a comparatively slight necrotic degree and area.In addition,the IVIS imaging revealed that PKH26-positive MenSCs were clearly retained in the liver initially and then diffused through the systemic circulation.Interestingly,the signal intensity in the liver increased obviously at 36 h,which corresponded to the biochemical result that liver function deteriorated most rapidly at 24-36 h.CONCLUSION Our study demonstrates the therapeutic efficacy and homing ability of transplanted MenSCs in a large animal model of ALF and suggests that MenSC transplantation could be a promising strategy for treating ALF.

Human umbilical cord blood-derived mesenchymal stem cells promote regeneration of crush-injured rat sciatic nerves

Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells （1 ~ 106） or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchvmal stem cells promote the functinnal r~.RcJv^rv nf P.n I^h-inillr^4 ~r^i~tit, n^r~e

Functional recovery and microenvironmental alterations in a rat model of spinal cord injury following human umbilical cord blood-derived mesenchymal stem cells transplantation

BACKGROUND： Transplantation of human umbilical cord blood-derived mesenchymal stem cells （MSCs） has been shown to benefit spinal cord injury （SCI） repair. However, mechanisms of microenvironmental regulation during differentiation of transplanted MSCs remain poorly understood. OBJECTIVE： To observe changes in nerve growth factor （NGF）, brain-derived neurotrophic factor （BDNF）, and interleukin-8 （IL-8） expression following transplantation of human umbilical cord-derived MSCs, and to explore the association between microenvironment and neural functional recovery following MSCs transplantation. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Department of Orthopedics, First Affiliated Hospital of Soochow University from April 2005 to March 2007. MATERIALS： Human cord blood samples were provided by the Department of Gynecology and Obstetrics, First Affiliated Hospital of Soochow University. Written informed consent was obtained. METHODS： A total of 62 Wister rats were randomly assigned to control （n = 18）, model （n = 22, SCI ＋ PBS）, and transplantation （n = 22, SCI ＋ MSCs） groups. The rat SCI model was established using the weight compression method. MSCs were isolated from human umbilical cord blood and cultured in vitro for several passages. 5-bromodeoxyuridine （BrdU）-Iabeled MSCs （24 hours before injection） were intravascularly transplanted. MAIN OUTCOME MEASURES： The rats were evaluated using the Basso, Beattie and Bresnahan （BBB） locomotor score and inclined plane tests. Transplanted cells were analyzed following immunohistochemistry. Enzyme-linked immunosorbant assay was performed to determine NGF, BDNF, and IL-8 levels prior to and after cell transplantation. RESULTS： A large number of BrdU-positive MSCs were observed in the SCI region of the transplantation group, and MSCs were evenly distributed in injured spinal cord tissue 1 week after transplantation. BBB score and inclined plane test results revealed significant functional improvement in the transplantation group compared to the model group （P 〈 0.05）, which was maintained for 2-3 weeks. Compared to the model group, NGF and BDNF levels were significantly increased in the injured region following MSCs transplantation at 3 weeks （P 〈 0.05）, but IL-8 levels remained unchanged （P 〉 0.05）. CONCLUSION： MSCs transplantation increased NGF and BDNF expression in injured spinal cord tissue. MSCs could promote neurological function recovery in SCI rats by upregulating NGF expression and improving regional microenvironments.

Differentiation of Menstrual Blood Derived Stem Cell (MensSCs) to Hepatocyte-Liked Cell on Three Dimensional Nanofiberscaffold: Poly Caprolacton (PCL)

Menstrual blood stem cells (MensSCs) have enormous potential as a source for cell replacement therapies. Since there is a major concern in utilization of nanofibers in tissue engineering of stem cells, we examined the potential of MensSCs to differentiate into hepatocytes, using different protocols and compare cells, with two-dimensional (2D) and three-dimensional (3D) culture systems. Cell characterization experiments of MensSCs have demonstrated that they are multipotent stem cells similar to mesenchymal stem cells, which can successfully differentiate into osteogenic and adipogenic lineages. The efficiency of the cells on the scaffold was appraised by scanning electron microscopy (SEM), MTT assay, and hematoxylin and eosin (H&E) staining. Thereafter, the differentiation protocols were developed by hepatocyte growth factor (HGF) and oncostatin M (OSM) with serum-supplemented or serum-free culture media up to 30 days. Immunofluorescence analysis and ELISA assay revealed the expression of albumin (ALB) in differentiated cells. Hepatocyte-like cells expressed liver-specific gene such as albumin(ALB), α-fetoprotein (AFP), tyrosine aminotransferase (TAT) and cytochrome P450 subunit 7a1 (Cyp7a1) at mRNA levels. In conclusion, the evidences presented in this study show that the nanofiber scaffold and MensSCs may provide a source of differentiated cells for treatment of liver diseases.

Human umbilical cord blood stem cells and brainderived neurotrophic factor for optic nerve injury: a biomechanical evaluation

Treatment for optic nerve injury by brain-derived neurotrophic factor or the transplantation of human umbilical cord blood stem cells has gained progress, but analysis by biomechanical indicators is rare. Rabbit models of optic nerve injury were established by a clamp. At 7 days after injury, the vitreous body received a one-time injection of 50 μg brain-derived neurotrophic factor or 1 × 10^6 human umbilical cord blood stem cells. After 30 days, the maximum load, maximum stress, maximum strain, elastic limit load, elastic limit stress, and elastic limit strain had clearly improved in rabbit models of optical nerve injury after treatment with brain-derived neurotrophic factor or human umbilical cord blood stem cells. The damage to the ultrastructure of the optic nerve had also been reduced. These findings suggest that human umbilical cord blood stem cells and brain-derived neurotrophic factor effectively repair the injured optical nerve, improve biomechanical properties, and contribute to the recovery after injury.

Human umbilical cord blood-derived stem cells and brain-derived neurotrophic factor protect injured optic nerve:viscoelasticity characterization

The optic nerve is a viscoelastic solid-like biomaterial.Its normal stress relaxation and creep properties enable the nerve to resist constant strain and protect it from injury.We hypothesized that stress relaxation and creep properties of the optic nerve change after injury.Moreover,human brain-derived neurotrophic factor or umbilical cord blood-derived stem cells may restore these changes to normal.To validate this hypothesis,a rabbit model of optic nerve injury was established using a clamp approach.At 7 days after injury,the vitreous body received a one-time injection of 50 μg human brain-derived neurotrophic factor or 1 × 106 human umbilical cord blood-derived stem cells.At 30 days after injury,stress relaxation and creep properties of the optic nerve that received treatment had recovered greatly,with pathological changes in the injured optic nerve also noticeably improved.These results suggest that human brain-derived neurotrophic factor or umbilical cord blood-derived stem cell intervention promotes viscoelasticity recovery of injured optic nerves,and thereby contributes to nerve recovery.

Safety of menstrual blood-derived stromal cell transplantation in treatment of intrauterine adhesion

BACKGROUND Intrauterine adhesion(IUA)can cause serious damage to women’s reproductive health,yet current treatment methods are difficult to achieve satisfactory results.In our previous studies,we demonstrated that menstrual-derived stromal stem cells(MenSCs),with high proliferative capacity and self-renewal ability,have a powerful therapeutic effect in patients with severe IUA.However,safety assessment of MenSCs transplantation is essential for its further application.AIM To evaluate the short-,medium-,and long-term biosafety of MenSCs via intrauterine transplantation in a rat model of IUA,with a focus on toxicity and tumorigenicity.METHODS MenSCs were injected into the sub-serosal layer of the uterus in an IUA rat model,for 3 d,3 mo,and 6 mo separately,to monitor the corresponding acute,sub-chronic,and chronic effects.Healthy rats of the same age served as negative controls.Toxicity effects were evaluated by body weight,organ weight,histopathology,hematology,and biochemistry tests.Tumorigenicity of MenSCs was investigated in Balb/c-nu mice in vivo and by colony formation assays in vitro.RESULTS Compared with the same week-old control group,all of the IUA rats receiving MenSC transplantation demonstrated no obvious changes in body weight,mainorgan weight,or blood cell composition during the acute,sub-chronic,and chronic observation periods.At the same time,serum biochemical tests showed no adverse effects on metabolism or liver and kidney function.After 4 wk of subcutaneous injection of Men SCs in Balb/c-nu nude mice,no tumor formation or cell metastasis was observed.Moreover,there was no tumor colony formation of Men SCs during soft agar culture in vitro.CONCLUSION There is no acute,sub-chronic,or chronic poisoning,infection,tumorigenesis,or endometriosis in rats with IUA after Men SC transplantation.The above results suggest that intrauterine transplantation of Men SCs is safe for endometrial treatment.

Transplantation of human umbilical cord blood mesenchymal stem cells to treat a rat model of traumatic brain injury

In the present study, human umbilical cord blood mesenchymal stem cells were injected into a rat model of traumatic brain injury via the tail vein. Results showed that 5-bromodeoxyuridine-labeled cells aggregated around the injury site, surviving up to 4 weeks post-transplantation. In addition, transplantation-related death did not occur, and neurological functions significantly improved. Histological detection revealed attenuated pathological injury in rat brain tissues following human umbilical cord blood mesenchymal stem cell transplantation. In addition, the number of apoptotic cells decreased. Immunohistochemistry and in situ hybridization showed increased expression of brain-derived neurotrophic factor, nerve growth factor, basic fibroblast growth factor, and vascular endothelial growth factor, along with increased microvessel density in surrounding areas of brain injury. Results demonstrated migration of transplanted human umbilical cord blood mesenchymal stem cells into the lesioned boundary zone of rats, as well as increased angiogenesis and expression of related neurotrophic factors in the lesioned boundary zone.

Umbilical cord blood mesenchymal stem cells protect amyloid-β42 neurotoxicity via paracrine

AIM:To understand the neuroprotective mechanism of human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSCs) against amyloid-β42(Aβ42) exposed rat primary neurons.METHODS:To evaluate the neuroprotective effect of hUCB-MSCs,the cells were co-cultured with Aβ42-exposed rat primary neuronal cells in a Transwell apparatus.To assess the involvement of soluble fac-tors released from hUCB-MSCs in neuroprotection,an antibody-based array using co-cultured media was conducted.The neuroprotective roles of the identified hUCB-MSC proteins was assessed by treating recombi-nant proteins or specific small interfering RNAs(siRNAs) for each candidate protein in a co-culture system.RESULTS:The hUCB-MSCs secreted elevated levels ofdecorin and progranulin when co-cultured with rat pri-mary neuronal cells exposed to Aβ42.Treatment with recombinant decorin and progranulin protected from Aβ42-neurotoxicity in vitro.In addition,siRNA-mediat-ed knock-down of decorin and progranulin production in hUCB-MSCs reduced the anti-apoptotic effects of hUCB-MSC in the co-culture system.CONCLUSION:Decorin and progranulin may be involved in anti-apoptotic activity of hUCB-MSCs exposed to Aβ42.

Research Progresses in the Inhibitory Effect of Bone Mesenchymal Stem Cell-Derived Exosome on Blood-Brain Barrier Disruption following Intracerebral Hemorrhage in Rats

Mesenchymal Stem Cells (MSCs) are a type of non-hematopoietic progenitor cells which have self-replication capacity and multilineage differentiation. They have widely applied in studies of various diseases due to their effects in damaged tissue repair, neuroprotection and immunoregulation. MSCs can secret exosomes through multiple ways in the physiological or pathological state. Many researches’ results on MSC-Exo show that it possesses many functions similar to MSCs, such as immunoregulation and regeneration promotion of damaged tissues. Hence, MSC-Exo is believed to have considerable research potentials in regenerative medicines. This study reviewed the research progresses on biological characteristics and functions of MSC-Exo.

Stimulation of decidua development by transplantation of endometrial stem cells

On all terms of pregnancy, insolvency of decidual reaction of endometrial cells is one of the reasons of miscarriages and fetal growth delay. The insufficient decidualization of endometrum leads to infertility in such pathologies, as Asherman’s syndrome and an endometrium atrophy. However, there are data on successful application of autologous bone marrow MSCs for Asherman’s syndrome treatment. The aim of this work was to assay the effect of endometrial mesenchymal stem cell (eMSC) transplantation for decidualization process in pseudopregnant rat. Our study showed that injection of human eMSC suspension into the uterine lumen of pseudopregnant rats facilitated more intensive development of decidua in comparison with phosphate buffed saline (PBS) injection in the control uterine horn. Histological analysis of decidua sections did not reveal any alterations in cell differentiation or tissue structure. In conclusion, we demonstrated for the first time that eMSC transplantation assists the development of all decidual tissue elements. It opens the possibility that eMSCs may be applied for cell therapy of infertility associated with decidualzation insufficiency.

Comparative analysis of biological characteristics of adult mesenchymal stem cells with different tissue origins

Objective: To invest the dif erences among mesenchymal stem cells(MSCs) derived from different tissues and their impacts on clinical applications. Methods: In this study, MSCs were isolated from adipose tissue(AD), umbilical cord tissue(UC), and menstrual blood(Men) and comparedtheir biological characteristics in terms of proliferation capacity, passage capacity, colony formation, and surface markers were compared. Results: The stem cells(SCs) obtained from different sources were all characterized as MSCs, but demonstrated some dif erences. Umbilical cord-derived MSCs(UCMSCs) were able to overcome density inhibition. The proliferation rate decreased in the order UCMSCs> Men SCs> ADSCs, while the colony-forming ability decreased in the order Men SCs> ADSCs> UCMSCs. Based on gene-expression data for MSCs from dif erent sources within the same donor, 768 Men SC genes were found that were specii cally upregulated or downregulated compared with bone marrow-derived MSCs and UCMSCs, most of which were involved in cell function-related pathways. In addition, Men SCs appeared to be superior in terms of immune inl ammation, stress response, and neural dif erentiation potentials, but weaker in terms of osteogenic and chondrogenic dif erentiation capacities, compared with UCMSCs and bone marrow-derived MSCs. Conclusions: Men SCs have higher extraction ei ciency, colony-forming ability, and long time passage capacity. Although the proliferation capacity is inferior to UCMSCs.

Stem cell transplantation for treating spinal cord injury A literature comparison between studies of stem cells obtained from various sources

OBJECTIVE： To identify global research trends of stem cell transplantation for treating spinal cord injury using a bibliometric analysis of the Web of Science. DATA RETRIEVAL： We performed a bibliometric analysis of data retrievals for stem cell transplantation for treating spinal cord injury from 2002 to 2011 using the Web of Science. SELECTION CRITERIA： Inclusion criteria： （a） peer-reviewed articles on stem cell transplantation for treating spinal cord injury that were published and indexed in the Web of Science; （b） type of articles： original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items; and （c） year of publication： 2002-2011. Exclusion criteria： （a） articles that required manual searching or telephone access; （b） documents that were not published in the public domain; and （c） a number of corrected papers from the total number of articles. MAIN OUTCOME MEASURES： （1） Annual publication output; （2） distribution according to country; （3） distribution according to institution; （4） distribution according to journals; （5） distribution according to funding agencies; and （6） top cited articles over the last 10 years. RESULTS： Bone marrow mesenchymal stem cells and embryonic stem cells have been widely used for treating spinal cord injury. In total, 191 studies of bone marrow mesenchymal stem cell transplantation and 236 studies of embryonic stem cell transplantation for treating spinal cord injury appeared in the Web of Science from 2002 to 2011, and almost half of which were derived from American or Japanese authors and institutes. The number of studies of stem cell transplantation for treating spinal cord injury has gradually increased over the past 10 years. Most papers on stem cell transplantation for treating spinal cord injury appeared in journals with a particular focus on stem cell research, such as Stem Cells and Cell Transplantation. Although umbilical cord blood stem cells and adipose-derived stem cells have been studied for treating spinal cord injury, the number of published papers was much smaller, with only 21 and 17 records, respectively, in the Web of Science. CONCLUSION： Based on our analysis of the literature and research trends, we found that stem cells transplantation obtained from various sources have been studied for treating spinal cord injury; however, it is difficult for researchers to reach a consensus on this theme.

8周运动预适应增强脂肪干细胞治疗心肌梗死大鼠的效果

背景:干细胞移植是心肌梗死的崭新疗法,然而梗死区域极其恶劣的微环境造成干细胞存活率低下并导致远期疗效甚微。运动预适应是一种通过运动诱导机体产生内源性保护效应的方式,可作为心脏康复预防与治疗的新策略。目的:评估运动预适应是否能够增强大鼠心肌梗死后脂肪干细胞移植的心脏保护效应,探讨血管生成在其中的作用机制。方法:6周龄雄性SD大鼠随机分为对照组、造模组、干细胞组以及干细胞运动组。利用冠状动脉闭塞术制作急性心肌梗死模型,对照组同期行假手术;干细胞运动组于造模前进行8周有氧运动,造模后30 min进行脂肪干细胞移植;干细胞组仅进行脂肪干细胞移植。干细胞移植后1 d和7 d,利用免疫印迹法测定心肌总Akt(t-Akt)、磷酸化Akt(p-Akt)、血管内皮生长因子(VEGF)、总内皮型一氧化氮合酶(t-eNOS)和磷酸化内皮型一氧化氮合酶(p-eNOS)蛋白表达量,计算p-Akt/t-Akt和p-eNOS/t-eNOS比值;4周后利用彩色多普勒超声诊断系统检测心脏结构与功能以及心肌血流量,TTC染色法检测心肌梗死面积,Masson染色法检测心肌间质胶原沉积,免疫荧光染色法测定心肌毛细血管密度,TUNEL染色法评估心肌细胞凋亡。结果与结论:(1)干细胞移植后4周:与对照组比较,造模组左心室缩短分数、左心室射血分数、心肌毛细血管密度和心肌血流量下降(P<0.05),心肌梗死面积、胶原容积分数和细胞凋亡增加(P<0.05);与造模组比较,干细胞组上述指标(除左心室缩短分数和左心室射血分数外)得到改善(P<0.05);与干细胞组比较,干细胞运动组以上各参数进一步改善(P<0.05)。(2)干细胞移植后1 d:与对照组比较,造模组t-Akt、p-Akt、VEGF、t-eNOS、p-eNOS蛋白表达量以及p-Akt/t-Akt、p-eNOS/t-eNOS比值均无显著性变化(P>0.05);与造模组比较,干细胞组上述指标均无显著性变化(P>0.05),干细胞运动组磷酸化p-Akt蛋白表达量以及p-Akt/t-Akt比值上调(P<0.05)。(3)干细胞移植后7 d:与对照组比较,造模组p-Akt、VEGF、p-eNOS蛋白表达量以及p-Akt/t-Akt、p-eNOS/t-eNOS比值下降(P<0.05);与造模组比较,干细胞组各参数均无显著性变化(P>0.05),干细胞运动组p-Akt、VEGF、p-eNOS蛋白表达量以及p-Akt/t-Akt、p-eNOS/t-eNOS比值升高(P<0.05)。结果表明:运动预适应可增强脂肪干细胞对心肌梗死大鼠心脏重塑的治疗效果,其机制与促进心肌血管生成并增加血流灌注有关。

Menstrual blood-derived mesenchymal stem cells differentiate into functional hepatocyte-like cells

Orthotopic liver transplantation(OLT)is the only proven effective treatment for both end-stage and metabolic liver diseases.Hepatocyte transplantation is a promising alternative for OLT,but the lack of available donor livers has hampered its clinical application.Hepatocyte-like cells(HLCs)differentiated from many multi-potential stem cells can help repair damaged liver tissue.Yet almost suitable cells currently identified for human use are difficult to harvest and involve invasive procedures.Recently,a novel mesenchymal stem cell derived from human menstrual blood(MenSC)has been discovered and obtained easily and repeatedly.In this study,we examined whether the MenSCs are able to differentiate into functional HLCs in vitro.After three weeks of incubation in hepatogenic differentiation medium containing hepatocyte growth factor(HGF),fibroblast growth factor-4(FGF-4),and oncostain M(OSM),cuboidal HLCs were observed,and cells also expressed hepatocyte-specific marker genes including albumin(ALB),α-fetoprotein(AFP),cytokeratin 18/19(CK18/19),and cytochrome P450 1A1/3A4(CYP1A1/3A4).Differentiated cells further demonstrated in vitro mature hepatocyte functions such as urea synthesis,glycogen storage,and indocyanine green(ICG)uptake.After intrasplenic transplantation into mice with 2/3 partial hepatectomy,the MenSC-derived HLCs were detected in recipient livers and expressed human ALB protein.We also showed that MenSC-derived HLC transplantation could restore the serum ALB level and significantly suppressed transaminase activity of liver injury animals.In conclusion,MenSCs may serve as an ideal,easily accessible source of material for tissue engineering and cell therapy of liver tissues.

Mini-review:Plasticity of human menstrual blood stem cells derived from the endometrium

Stem cells can be obtained from women's menstrual blood derived from the endometrium.The cells display stem cell markers such as Oct-4,SSEA-4,Nanog,and c-kit(CD117),and have the potent ability to differentiate into various cell types,including the heart,nerve,bone,cartilage,and fat.There has been no evidence of teratoma,ectopic formation,or any immune response after transplantation into an animal model.These cells quickly regenerate after menstruation and secrete many growth factors to display recurrent angiogenesis.The plasticity and safety of the acquired cells have been demonstrated in many studies.Menstrual blood-derived stem cells(MenSCs) provide an alternative source of adult stem cells for research and application in regenerative medicine.Here we summarize the multipotent properties and the plasticities of MenSCs and other endometrial stem cells from recent studies conducted both in vitro and in vivo.

Human adult stem cells from menstrual blood and endometrial tissue

The article ＂Plasticity of human menstrual blood stem cells derived from the endometrium＂ by Lin et al （2011）, published in Journal of Zhejiang University- SCIENCE B （Biomedicine ＆ Biotechnology）, described a newly identified mesenchymal-like stem cell （MSC） from human menstrual blood known as MenSC. Here we describe the latest findings in this area and clarify the difference between human adult stem cells from menstrual blood and endometrial tissue.

黑枸杞花青素联合人脂肪源性血管外膜细胞支持脐血造血干/祖细胞的增殖

背景:黑枸杞花青素(Anthocyanins in Lycium ruthenicum Murr,ALRM)是黑枸杞中重要的活性成分之一,具有抗氧化、免疫调节等功效。人脂肪源性血管外膜细胞(CD146^(+)hAD-PCs)是骨髓间充质干细胞的前体细胞,在体外具有促进造血干/祖细胞增殖与分化的功能。ALRM联合CD146^(+)hAD-PCs对脐血造血干/组细胞的体外支持作用有待于研究。目的:探讨ALRM联合CD146^(+)hAD-PCs对脐血CD34^(+)造血干/祖细胞体外扩增的支持作用。方法:CCK-8法检测不同质量浓度ALRM(0,200,400,600,800,1000 mg/L)对CD146^(+)hAD-PCs增殖的影响;流式细胞术检测ALRM对CD146^(+)hAD-PCs细胞周期的影响。共培养实验分为空白组、ALRM组、CD146^(+)hAD-PCs组、ALRM+CD146^(+)hAD-PCs组,分析ALRM联合CD146^(+)hAD-PCs对脐血CD34^(+)造血干/祖细胞的体外支持作用。共培养1,2,4周,比较扩增后细胞数量、集落形成单位数量,流式细胞仪检测细胞免疫表型,ELISA检测细胞因子水平。结果与结论:(1)ALRM质量浓度为200 mg/L时,CD146^(+)hAD-PCs活力最高,CD146^(+)hAD-PCs的G_(0)/G_(1)期细胞比例下降,S期、G_(2)/M期细胞比例上升(P<0.01)。(2)脐血CD34^(+)造血干/祖细胞数量变化:在共培养1,2,4周时ALRM+CD146^(+)hAD-PCs组高于ALRM组(P均<0.05),在共培养2,4周时ALRM+CD146^(+)hAD-PCs组高于CD146^(+)hAD-PCs组(P均<0.05),ALRM组与空白组随着共培养时间延长细胞数量逐渐减少。(3)集落形成能力及免疫表型分析:在共培养1,2周时ALRM+CD146^(+)hAD-PCs组的集落形成单位数量高于CD146^(+)hAD-PCs组和ALRM组(P均<0.05);在共培养1,2,4周时ALRM+CD146^(+)hAD-PCs组CD45^(+)、CD34^(+)CD33^(-)细胞比例高于CD146^(+)hAD-PCs组(P均<0.01)。(4)细胞因子变化:在共培养4周时ALRM+CD146^(+)hAD-PCs组的白细胞介素2水平高于ALRM组、CD146^(+)hAD-PCs组(P<0.05);在共培养2,4周时ALRM+CD146^(+)hAD-PCs组白细胞介素3水平高于CD146^(+)hAD-PCs组(P<0.05);在共培养1周时ALRM+CD146^(+)h AD-PCs组的粒细胞集落刺激因子水平高于CD146^(+)hAD-PCs组,在共培养2周时高于ALRM组、CD146^(+)hAD-PCs组(P<0.01);在共培养1,2,4周时ALRM组、ALRM+CD146^(+)hAD-PCs组的干扰素γ水平低于CD146^(+)hAD-PCs组(P<0.05)。(5)由于空白组无基质细胞,脐血CD34^(+)造血干/祖细胞在共培养1周之后就无法计数,未进行免疫表型、集落分析和细胞因子检测。(6)结果表明:ALRM可以通过促进CD146^(+)hAD-PCs增殖和细胞周期转化进而促进脐血CD34^(+)造血干/祖细胞的体外扩增,在造血干细胞移植研究方面具有重要价值。

Proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood under the culture conditions of hypoxia and serum deprivation

Background The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood （PB-MSCs） were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair. Methods MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor （G-CSF） and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension （20% 02 and 2% O2）, with or without 10% fetal bovine serum （FBS） conditions： 20% 02 and 10% FBS complete medium （normal medium, N）, 20% 02 and serum deprivation medium （D）, 2% 02 and 10% FBS complete medium （hypoxia, H）, 2% 02 and serum deprivation （HD）. Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling （TUNEL） staining. Results Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow （BM-MSCs）, and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II （MHC II）. They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia. Conclusions PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.