Hupo powder promotes autophagy of menstrual blood-derived stem cells from patients with endometriosis

Objective:To explore the effect of Hupo powder(HP)on autophagy in menstrual blood-derived stem cells(MenSCs)with endometriosis(EMT).Methods:EMT MenSCs(E-MenSCs)and healthy MenSCs(H-MenSCs)were isolated from the menstrual blood of patients with EMT and healthy female participants,respectively.We identified their stem cells’characteristics via adipogenic and osteogenic differentiation.Twelve male SpragueeDawley rats received 0.9% NaCl and HP-dispensing granules by gastric irrigation to prepare blank serum and medicated serum,respectively.We used serum concentrations of 5%,10%,and 20%,each at administered times of 12,24,and 48 h to select the best condition.These cells were divided into three groups:blank serum of the control group,blank serum of the model group,and medicated serum of the HP group.H-MenSCs were used in the control group,while E-MenSCs were used in the model and HP groups.We analyzed cell viability using a cell counting kit-8 assay,observed cell morphology,evaluated the amounts of auto-phagosomes and autolysosomes by transmission electron microscopy,and detected the protein expression of autophagy markers(LC3-II and Beclin1)by Western blot.Results:E-MenSCs and H-MenSCs became long fusiform with a diffuse radial pattern,forming lipid droplets and calcium nodules after adipogenic and osteogenic differentiation.We then used the best conditiond 20% serum and 48 hdfor the subsequent experiments.In contrast to the model group,the HP group exhibited lower cell viability(=0.007),larger amounts of autophagosomes and autolysosomes(P<0.001 and P=0.001,respectively),and higher expression of LC3-II and Beclin1(P=0.021 and P=0.019,respectively).Conclusion:Hupo powder can promote autophagy in E-MenSCs,which might be one of the mechanisms underlying its therapeutic effects.

Human menstrual blood-derived stem cells as immunoregulatory therapy in COVID-19:A case report and review of the literature

BACKGROUND The coronavirus disease 2019(COVID-19)caused by novel coronavirus 2019 in December 2019 has spread all around the globe and has caused a pandemic.There is still no current effective guidance on the clinical management of COVID-19.Mesenchymal stem cell therapy has been shown to be one of the therapeutic approaches to alleviate pneumonia and symptoms through their immunomodulatory effect in COVID-19 patients.CASE SUMMARY We describe the first confirmed case of COVID-19 in Hangzhou to explore the role of human menstrual blood-derived stem cells(MenSCs)in the treatment of COVID-19.Moreover,we review the immunomodulation effect including nonspecific and specific immune functions of MenSCs for the therapy of COVID-19.CONCLUSION MenSCs can be helpful to find a promising therapeutic approach for COVID-19.

Human menstrual blood-derived stem cells alleviate autoimmune hepatitis via JNK/MAPK signaling pathway in vivo and in vitro

Autoimmune hepatitis(AIH)is a severe globally distributed liver disease that could occur at any age.Human menstrual blood-derived stem cells(MenSCs)have shown therapeutic effect in acute lung injury and liver failure.However,their role in the curative effect of AIH remains unclear.Here,a classic AIH mouse model was constructed through intravenous injection with concanavalin A(Con A).MenSCs were intravenously injected while Con A injection in the treatment groups.The results showed that the mortality by Con A injection was significantly decreased by MenSCs treatment and liver function tests and histological analysis were also ameliorated.The results of phosphoproteomic analysis and RNA-seq revealed that MenSCs improved AIH,mainly by apoptosis and c-Jun N-terminal kinase/mitogen-activated protein signaling pathways.Apoptosis analysis demonstrated that the protein expression of cleaved caspase 3 was increased by Con A injection and reduced by MenSCs transplantation,consistent with the TUNEL staining results.An AML12 co-culture system and JNK inhibitor(SP600125)were used to verify the JNK/MAPK and apoptosis signaling pathways.These findings suggested that MenSCs could be a promising strategy for AIH.

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.

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.

High tibial osteotomy with human umbilical cord blood-derived mesenchymal stem cells implantation for knee cartilage regeneration

BACKGROUND High tibial osteotomy(HTO)is a well-established method for the treatment of medial compartment osteoarthritis of the knee with varus deformity.However,HTO alone cannot adequately repair the arthritic joint,necessitating cartilage regeneration therapy.Cartilage regeneration procedures with concomitant HTO are used to improve the clinical outcome in patients with varus deformity.AIM To evaluate cartilage regeneration after implantation of allogenic human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSCs)with concomitant HTO.METHODS Data for patients who underwent implantation of hUCB-MSCs with concomitant HTO were evaluated.The patients included in this study were over 40 years old,had a varus deformity of more than 5°,and a full-thickness International Cartilage Repair Society(ICRS)grade IV articular cartilage lesion of more than 4 cm2 in the medial compartment of the knee.All patients underwent second-look arthroscopy during hardware removal.Cartilage regeneration was evaluated macroscopically using the ICRS grading system in second-look arthroscopy.We also assessed the effects of patient characteristics,such as trochlear lesions,age,and lesion size,using patient medical records.RESULTS A total of 125 patients were included in the study,with an average age of 58.3±6.8 years(range:43-74 years old);95(76%)were female and 30(24%)were male.The average hip-knee-ankle(HKA)angle for measuring varus deformity was 7.6°±2.4°(range:5.0-14.2°).In second-look arthroscopy,the status of medial femoral condyle(MFC)cartilage was as follows:73(58.4%)patients with ICRS grade I,37(29.6%)with ICRS grade II,and 15(12%)with ICRS grade III.No patients were staged with ICRS grade IV.Additionally,the scores[except International Knee Documentation Committee(IKDC)at 1 year]of the ICRS grade I group improved more significantly than those of the ICRS grade II and III groups.CONCLUSION Implantation of hUCB-MSCs with concomitant HTO is an effective treatment for patients with medial compartment osteoarthritis and varus deformity.Regeneration of cartilage improves the clinical outcomes for the patients.

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.

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.

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.

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.

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.

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.

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.

经血间充质干细胞外泌体对人卵巢癌细胞A2780生物学行为的影响

目的 探讨经血源性间充质干细胞(menstrual blood-derived mesenchymal stem cells,MenSCs)来源的外泌体(MenSCs-derived exosomes,MenSCs-Exos)对人卵巢癌细胞A2780增殖、迁移、侵袭能力和细胞周期分布的影响和机制。方法 分离纯化MenSCs,流式细胞术鉴定其细胞表型,油红O染色观察其成脂分化能力;超速离心法分离MenSCs-Exos,电子显微镜观察外泌体形态,纳米颗粒追踪分析其大小及分布,蛋白免疫印迹法检测外泌体标志性蛋白分子CD9、TSG101、calnexin的表达。采用CCK-8实验、划痕实验、Transwell细胞体外侵袭实验、流式细胞仪检测MenSCs-Exos对A2780细胞增殖、迁移、侵袭能力和细胞周期分布的影响;蛋白免疫印迹法检测Wnt/β-catenin信号通路相关蛋白和迁移相关蛋白的表达。结果 成功分离具有干细胞表型和成脂分化潜能的MenSCs,同时检测发现MenSCs-Exos为边缘清晰、大小分布均匀的膜性囊泡,粒径为30~150 nm,平均为72.89 nm,膜表面标志性蛋白CD9、TSG101表达呈阳性,calnexin表达呈阴性。与对照组相比,MenSCs-Exos能增强A2780细胞增殖、迁移和侵袭能力,影响其细胞周期分布,并且可上调β-catenin、c-Myc、TCF4、Cyclin D1、Vimentin、N-cadherin的表达,下调E-cadherin的表达。结论 MenSCs-Exos可能是通过激活Wnt/β-catenin信号通路促进A2780细胞增殖,并通过下调E-cadherin的表达促进A2780细胞的迁移和侵袭。

月经血源性间充质干细胞来源外泌体对SD大鼠皮肤创面愈合的促进作用及其机制

目的观察月经血源性间充质干细胞(MenSCs)中分离的外泌体(MenSC-Exos)对SD大鼠皮肤创面愈合的促进作用,并探讨其机制。方法分离培养健康成年女性志愿者的MenSCs,提取MenSC-Exos;将SD大鼠随机分为实验组和对照组,制作背部皮肤机械创面,分别给予MenSC-Exos或PBS治疗;观察创面愈合情况并计算创面愈合率,采用HE染色观察创面炎症细胞浸润情况,免疫荧光法检测M1、M2型巨噬细胞,采用免疫组化法检测创面组织中的白细胞介素(IL)-6、IL-10、新生血管、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白,采用Masson染色检测创面胶原蛋白含量及成熟度。培养成纤维细胞并分成外泌体组和PBS组,分别加入MenSC-Exos或PBS,采用CCK-8法检测细胞增殖能力,划痕实验检测细胞迁移能力。结果实验组术后第7、14天创面愈合率、Ⅲ型胶原蛋白沉积率和胶原蛋白含量高于对照组,Ⅰ型胶原蛋白沉积率低于对照组(P均<0.05)。实验组术后第7天创面炎症细胞数量和M1/M2低于对照组,M2型巨噬细胞数量高于对照组(P均<0.05);与对照组相比,实验组术后第3、7天创面IL-6表达降低,IL-10表达及血管内皮细胞表达升高(P均<0.05)。外泌体组培养1~5 d细胞增殖能力均高于PBS组,外泌体组培养24 h细胞迁移率高于PBS组(P均<0.05)。结论MenSC-Exos能够促进大鼠皮肤创面愈合,机制可能与调节炎症反应、促进血管生成和成纤维细胞增殖、调节胶原重塑有关。

经血源子宫内膜干细胞移植通过免疫调节缓解化疗引起的小鼠肠道损伤和菌群失调

目的:研究经血源子宫内膜干细胞(menstrual blood-derived endometrial stem cells,MenSCs)对化疗引起的小鼠肠道黏膜炎和菌群失调的治疗效果及潜在机制。方法:将小鼠随机分为正常组、顺铂(cisplatin,Cis)组和Cis+MenSC组,每组10只:Cis组和Cis+MenSC组小鼠采用腹腔注射Cis(2 mg·kg^(−1)·d^(−1),连续5 d)的方式建立小鼠化疗性肠黏膜炎模型,正常组小鼠经腹腔注射同等体积生理盐水;Cis+MenSC组小鼠经尾静脉移植MenSCs,而正常组与Cis组小鼠经尾静脉注射同等体积生理盐水。给药过程中观察小鼠体征及体重变化,实验结束后分离小鼠小肠、结肠部位,通过HE染色评估小鼠肠道组织形态学改变;免疫组化染色检测肠道组织中F4/80和IL-6阳性细胞率;Western blot检测肠道组织中肠道紧密连接蛋白、炎症因子及凋亡相关蛋白的变化;粪便微生物菌群16S rDNA扩增子测序检测小鼠肠道微生物群多样性和丰度的改变情况。结果:与Cis组小鼠相比,MenSCs移植组小鼠体重显著增加,HE染色显示小肠及结肠组织炎性细胞浸润减少,肠绒毛水肿减轻,腺体排列较有序;Western blot结果显示MenSCs移植可显著上调Cis损伤后小鼠肠道组织中紧密连接蛋白ZO-1和occludin的表达(P<0.05)。随后,免疫组化染色和Western blot结果显示MenSCs移植可显著降低Cis损伤后小鼠肠道组织中巨噬细胞数量(P<0.01),下调肠道组织中促炎因子IL-1β和IL-6和促凋亡蛋白Bax的表达(P<0.01);同时显著上调肠道组织中抑炎因子IL-10和抗凋亡相关蛋白Bcl-2的表达(P<0.01)。进一步粪便微生物测序结果显示,MenSCs移植可改善Cis诱导的肠道菌群失调,显著降低有害微生物艾森伯格氏菌(Eisenbergiella tayi)和人结肠厌氧棍状菌(Anaerotruncus colihominis)的丰度(P<0.01),同时显著增加肠道中有益微生物乳杆菌(Lactobacillus apodemi)的丰度(P<0.05),有助于恢复健康肠道菌群结构和功能。结论:MenSCs移植能够减轻化疗药物造成的小鼠肠道屏障损伤,减轻化疗相关小鼠黏膜炎的症状,有助于恢复肠道菌群稳态。