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 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.

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 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.

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.

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.

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.

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.

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.

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。

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.

经血间充质干细胞及其培养液对化疗致早发性卵巢功能不全小鼠的治疗作用

目的探讨经血间充质干细胞(MenSCs)及其培养液(MenSCs-CM)对化疗致早发性卵巢功能不全(POI)小鼠的治疗作用。方法将健康的雌性ICR小鼠分为4组:对照组、POI组、POI+MenSCs组和POI+MenSCs-CM组。POI组、POI+MenSCs组和POI+MenSCs-CM组用环磷酰胺(CTX)建立POI模型,建模成功后分别进行对应治疗处理。测量小鼠卵巢脏器系数,HE染色检查卵巢卵泡情况,ELISA测定血清卵泡刺激素(FSH)和雌激素(E 2)水平,TUNEL法观察卵巢细胞凋亡情况,交配实验检测生育力,并对4组进行比较。结果治疗第7天和第14天,POI组的卵巢脏器系数均显著低于对照组(P<0.05),POI+MenSCs组和POI+MenSCs-CM组均显著高于POI组(P<0.05);治疗第7天和第14天,POI组的血清FSH水平显著高于对照组(P<0.05),POI+MenSCs组和POI+MenSCs-CM组均显著低于POI组(P<0.05);治疗第14天,POI+MenSCs组和POI+MenSCs-CM组的血清E 2水平均显著高于POI组(P<0.05)。POI+MenSCs组和POI+MenSCs-CM组的原始卵泡、初级卵泡、次级卵泡和窦状卵泡数量均显著多于POI组(P<0.05),闭锁卵泡数显著少于POI组(P<0.05)。TUNEL法结果表明,凋亡细胞大部分为成熟卵泡周边及内部的颗粒细胞(GCs),POI+MenSCs组和POI+MenSCs-CM组的凋亡指数均显著小于POI组(P<0.05),且两组的产仔数量和幼崽体重均显著高于POI组(P<0.05)。结论来源于经血的MenSCs及其培养液可有效改善POI小鼠的卵巢功能,并可提升其生育能力。

经血来源干细胞及其外泌体的应用现状和前景

背景:经血来源干细胞是从女性月经血中提取的,有多向分化潜能的间充质干细胞,具有易获取、弱免疫原性、增殖能力强、传代稳定及无伦理争议等特点。经血来源干细胞中提取的外泌体在组织再生和损伤恢复方面具有与经血来源干细胞相同的功效。经血来源干细胞及其外泌体已经在多种疾病治疗的临床前研究和临床研究中取得了令人兴奋的成果。目的:文章将就经血来源干细胞及其外泌体相关的基础研究与临床应用研究进展及未来潜在的应用前景和需要考虑的问题加以综述。方法:应用计算机检索PubMed、中国知网期刊全文数据库2000-2022年与经血来源干细胞和经血来源干细胞外泌体相关的文献,最终筛选出75篇进行综述分析。结果与结论:①经血来源干细胞是具有多向分化功能的间充质干细胞,具有来源丰富、免疫原性弱和增殖能力强的特点。②经血来源干细胞外泌体是发挥经血来源干细胞旁分泌功能的主要介质,在促进组织再生和损伤修复方面具有与来源细胞相同的功效。二者都具有抗炎、抗凋亡和促进组织器官功能恢复的能力,并在各系统疾病的动物实验模型中展现了较为优秀的治疗水平,其中经血来源干细胞已经在宫腔粘连、充血性心力衰竭、多发性硬化及病毒性肺炎等疾病中应用于临床患者,在没有产生肿瘤或过敏反应等不良作用的情况下显著延缓了疾病的进展。③但从经血来源干细胞从2007年被发现至今只过了短短15年,因此经血来源干细胞及其外泌体具有试验样本不足和应用年限过短的短板,仍需长期的临床前研究验证其安全性和有效性,此外,要真正应用于临床,也需要找到更加规范的给药方式,因此未来出台相关应用指南也是必要的。④经血来源干细胞及其外泌体结合三维支架技术已在盆腔器官脱垂、糖尿病导致的皮肤损伤、骨软骨缺损及膀胱壁重建的动物实验研究中验证了治疗的有效性,未来希望借助经血来源干细胞及其外泌体的各种优点与组织工程学和材料学相结合,逐步开展临床研究。

经血源子宫内膜干细胞复合3D打印PLGA支架体外培养的相容性研究

[目的]通过经血源子宫内膜干细胞(menstrual blood-derived mesenchymal stem cells,Men SCs)与3D打印PLGA支架材料的复合培养,探讨构建组织工程化骨软骨的可行性。[方法]预处理3D打印PLGA支架,取第5代Men SCs,倒置显微镜下观察细胞生物学特性,按1.0×106/m L的密度接种到PLGA支架材料上复合培养,通过倒置显微镜、Mico-CT、扫描电镜和组织病理学进行观察,并借此判断Men SCs与3D打印PLGA支架材料复合体外培养的融合性。[结果]Men SCs生长良好,细胞平铺生长,呈梭形或纺锤形,胞质薄,核圆居中,具有成纤维细胞形态。细胞传至第5代,为长梭形细胞单层,呈漩涡状排列。PLGA支架的微观呈现相互连通的多孔结构,结构疏松,孔隙大且互相交通,孔壁较薄。Men SCs在PLGA支架材料上生长状态良好,细胞在支架表面和内部生长,支架孔径内有细胞基质样组织连接,表明Men SCs与3D打印PLGA支架材料复合体外培养的融合性良好。[结论]经血源Men SCs是骨组织工程适宜的种子细胞,与3D打印的PLGA支架材料复合可体外构建组织工程骨。

人经血来源间充质干细胞对小鼠A549肺癌化疗效果的影响研究

目的：探讨人经血来源间充质干细胞（mesenchymal stem cells，MenSCs）对小鼠A549肺癌化疗效果的影响。方法：取20只BALB/C裸小鼠建立肺癌模型，随机分为实验组和对照组各10只；对照组仅给予化疗干预，实验组则于化疗期间将某机构赠予的MenSCs经DiI荧光标记后于鼠尾静脉（3＆#215;105/只）进行注射。肺癌造模后14 d，两组小鼠均断颈椎处死并收集其肿瘤，比较两组小鼠瘤体体积、瘤体质量、抑瘤率及双肺湿重、肿瘤转移率、肿瘤转移个数、抑制转移率差异。结果：20只BALB/C裸小鼠均进行统计，无脱落报告。20只小鼠接瘤后第7天在右前肢腋下均可触及瘤体长出，两组小鼠瘤体生长曲线比较，差异无统计学意义（P〉0.05）。两组小鼠的平均瘤体体积和平均瘤体质量比较，差异均无统计学意义（P均〉0.05）；抑瘤率为8.4%。实验组小鼠双肺湿重、肿瘤转移率及肿瘤转移个数均显著低于对照组小鼠，差异有显著性（P〈0.05）；抑制转移率为71.8%。结论：MenSCs协同化疗在抑制小鼠肺癌肿瘤转移方面有一定效果。

经血源性间充质干细胞的分离培养与鉴定

目的建立分离培养经血源性间充质干细胞(MenSCs)的方法。方法通过Ficol密度梯度离心法从经血中分离MenSCs,培养后观察细胞形态特征和生长特性,绘制生长曲线。采用流式细胞仪栓测细胞表面抗原标志物的表达,并检测其体外成脂分化的潜能。结果经Fioll密度梯度离心法可以分离到贴壁生长,高表达CD73、CD90和CD105,低表达CD14、CD34、CD45及CD19,符合间充质干细胞(MSCs)表面分子标志物表达标准的呈纺锤状的MSCs。细胞成脂分化诱导后可以得到脂滴细胞。结论应用Ficoll密度梯度离心法可从经血中分离到MSCs,即MenSCs。

氯化锂对经血源性间充质干细胞增殖和神经分化的影响

目的:探讨Wnt/β-catenin通路激活剂氯化锂(Li Cl)对经血源性间充质干细胞(MMCs)增殖和神经分化的影响。方法:CCK-8法检测不同浓度(0、4、10、20、40 mmol/L)Li Cl作用4 d对MMCs增殖的影响,采用半定量PCR和Western blot法检测4 mmol/L Li Cl对MMCs中β-catenin表达的影响;采用免疫细胞化学检测对照组、诱导分化组(DMSO和β-ME联合诱导分化)和4 mmol/L Li Cl诱导组细胞中NF和NSE阳性细胞率,采用半定量PCR检测NF和NSE在转录水平的表达。结果:低浓度(4、10 mmol/L)Li Cl促进MMCs增殖,高浓度(20、40 mmol/L)Li Cl抑制MMCs增殖。与对照组相比,4 mmol/L Li Cl处理可增加β-catenin蛋白的表达水平(t=64.500,P<0.001),而mRNA的表达水平无明显变化(t=1.606,P=0.183)。诱导分化组细胞中NF和NSE mRNA的表达水平及蛋白的阳性表达率升高(P<0.05);与诱导分化组相比,4 mmol/L Li Cl诱导组NF和NSE mRNA的表达水平及蛋白的阳性表达率降低(P<0.05)。结论:4 mmol/L Li Cl可通过增强β-catenin蛋白的表达激活Wnt/β-catenin信号通路,促进MMCs的增殖;而在诱导分化过程中,Li Cl通过抑制NF和NSE在转录和蛋白水平的表达抑制MMCs向神经元分化。