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Germline cells derived from mesenchymal stem cells, with the focus on Wharton's jelly

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Asian pacific Journal of Reproduction 2018年第2期7卷 49-55页

作者： Hossein Yazdekhasti Jalil Hosseini Zahra Rajabi Maryam Hosseinzadeh Shirzeyli Fereshte Aliakbari Department of Cell Biology Center for Research in Contraceptive and Reproductive HealthUniversity of VirginiaCharlottesvilleVA22908USA Infertility and Reproductive Health Research Center Shahid Beheshti University of Medical SciencesTehranIran Department of Anatomy School of MedicineTehran University of Medical SciencesTehranIran Department of Biomedical Engineering University of VirginiaCharlottesvilleVA22908USA Department of Biology and Anatomical Sciences School of MedicineShahid Beheshti University of Medical SciencesTehranIran

Previous attempts have indicated that mesenchymal stem cells (MSCs) are a valuable source and candidate and new approach for tissue engineering and reproductive medicine. MSCs have this potential to be induced and differentiated in an appropriatein vivoandin vitro condition toward various cell lineages and then they can be applied in cell therapies and clinical applications. During recent two decades, various sources have demonstrated they are a great source for MSCs, including bone marrow, the human umbilical cord as well as Wharton's jelly. Due to discarding after birth, easily accessible cells and less ethical concerns, these cells have attracted more and more scientists' attention. Infertility and reproduction diseases have provided special opportunity to examine the efficiency of MSCs in this kind of application. Based on recent investigations, MSCs embedded in Wharton's jelly tissue are more appealing for cell therapies, especially in infertility treatment purposes. So, differentiation of MSCs embedded in Wharton's jelly tissue into germ layer cells for cell-based therapy purposes is now under intensive study.

关键词： Mesenchymal stem cell Bone marrow stem cell umbilical cord Wharton's jelly

Inhibition Mechanism of Novel Pyrazolo[1,5-a]pyrazin-4(5H)-one Derivatives Against Proliferation of A549 and H322 Cancer Cells

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Chinese Medical Sciences Journal 2015年第4期30卷 260-265页

作者： Jin-hui Shao Gui-hua Feng Department of Medical Morphology School of MedicineHubei University of Art and Science Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology School of Life Science Shandong University

Objective To explore the inhibition mechanism and safety of pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives against proliferation of human lung cancer A549 cells, H322 cells, and human umbilical vein endothelial cell(HUVEC). Methods Cells were treated with 40 μmol/L of the ppo3 a, ppo3 b, ppo3 i, and 0.1% DMSO(control) for 48 hours, respectively. Apoptosis was determined by Hoechst 33258 staining assay in H322 and A549 cells. Cell cycle distribution was determined by flow cytometry analysis in A549 cell. LC3-II, p53, and heat shock protein(HSP) 70 protein levels were detected by Western blotting in A549 cells treated with ppo3 b for 48 hours. The morphology and viability of HUVEC were observed by inverted microscope and sulforhodamine B(SRB) assay. Results Ppo3 a, ppo3 b, and ppo3 i significantly induced apoptosis in H322 and A549 cells. A strong G1-phase arrest was concomitant with the growth inhibitory effect on A549 cells. Ppo3 b effectively elevated the p53 protein level, but significantly reduced the HSP70 protein level. There were no significantly inhibitory effect on the morphology and viability of HUVEC when treated with ppo3 a, ppo3 b, and ppo3 i. Conclusions ppo3 a, ppo3 b, and ppo3 i could inhibit H322 proliferation through apoptosis and inhibit A549 through apoptosis and G1-phase arrest. The protein p53 and HSP70 might involve in the inhibition effects. These derivatives might be a clue to find effective and safe drug for lung cancers.

关键词： pyrazole apoptosis lung cancer cell p5 3 human umbilical vein endothelial cell

The protective effect and action mechanisms of Icariin on oxidative injury of HUVEC induced by H2 02

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中国药理学通报 2015年第B11期31卷 127-128页

作者： LUOYun-mei FUXiao-xia LILi-sheng DepartmentofPharmacology ZunyiMedicalCollege KeyLaboratoryforBasicPharmacologyofMinistryofEducation ZunyiMedicalCollegeZunyiGuizhou563006

Aim To investigate the protective effects of icariin（ICA） on oxidative injury of human umbilical vascu- lar endothelial cells （HUVEC） caused by H202 and analysis action machenisms of ICA. Methods HUVEC were cultured in DMEM medium with 10% fetal bovine serum and 1% antibiotic mixture comprising penicillin and strep- tomycin, at 37℃; with 5% CO2 in vitro. The model of oxidative injury was established by H202 （ 750 μmol · L^-1 ）. The cells were divided into six groups： control group, H202 group, H202 ＋ ICA（ 10-8, 10-7, 10-6 tool · L^-1 ） group and H202 ＋ DMSO （solvent） ; HUVEC were treated with different concentration ICA according to grou- os before 12h H202 treatment and cultured for 12, 24 and 36h respectively, the cell vitality （OD value） and the level of ROS was measured by MTT and ELISA. The apoptosis and death of HUVEC were detected using Annexin V-FITC/PI kit. Real Time RT-PCR was used to detect Bcl-2 and Bax mRNA expression in the HUVEC. Results Compared with control group, cell vitality significantly declined （P 〈 0.01 ） , the level of ROS and the number of apoptotic HUVEC notably increased （P 〈 0. 01 ） in H202 group. Compared with H202 group, ICA treatment could increase HUVEC vitality and reduce level of ROS （P 〈 0.01 or P 〈 0. 05） with manners of time-dependence and dose-dependence,ICA treatment could decrease the apoptosis and death rate of HUVEC （P 〈 0.01 ）. ICA treat- ment could also down-regulate the expression of Bax mRNA （ P 〈 0.05 ） and up-regulate the expression of Bcl-2 （ P 〈 0.05）. Conclusion The ICA can alleviate the oxidative injury of HUVEC induced by H202 treatment. The ac- tion mechanisms of ICA may, at least partly, be related to inhibit HUVEC apoptosis caused by H202 treatment through up-regulation of expression of Bcl-2 and down-regulation of expression of Bax mRNA.

关键词： ieariin human umbilical vascular endothelial cells ( HUVEC) hydrogen peroxide ( H202) oxida-tive injury

人脐带血间充质干细胞移植对急性心肌梗死大鼠心功能的影响

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血栓与止血学 2010年第6期16卷 244-249页

作者：何志裕陆东风广州医学院第一附属医院心内科广州510120

目的研究脐带血来源的间充质干细胞(mesenchymal stem cells,MSCs)体外分离、纯化及培养的条件,以建立稳定的体外培养体系,满足实验和临床的需要;探讨经尾静脉移植脐血间充质干细胞对心肌梗死大鼠心功能的影响。方法无菌条件下采集健康... 详细信息

目的研究脐带血来源的间充质干细胞(mesenchymal stem cells,MSCs)体外分离、纯化及培养的条件,以建立稳定的体外培养体系,满足实验和临床的需要;探讨经尾静脉移植脐血间充质干细胞对心肌梗死大鼠心功能的影响。方法无菌条件下采集健康育龄产妇正常分娩胎儿的脐带血42份,脐血随机分成3组:FBS(胎牛血清)未包被组、FBS包被组和mesencult培养基组。FBS未包被组和FBS包被组均使用含10%FBS的LG-DMEM培养基、mesencult培养基组使用专用来培养干细胞的mesencult培养基,与其添加物配合使用。3组均用Ficoll淋巴细胞分离液,密度梯度法分离脐血单个核细胞(MNC),将单个核细胞按密度1×108/ml接种于T25培养瓶中,置37℃饱和湿度含5%CO2孵育箱培养,72 h后全量换液,去除未贴壁细胞,以后均7 d换液1次。待细胞长到80%铺满瓶底时结束原代培养。按1∶1进行消化传代。观察不同培养条件对脐血MSCs生长的影响。取P2代细胞用流式细胞仪检测细胞表面CD 29、CD 34、CD45、CD 105标志。将36只SD大鼠随机分成MSCs移植组、假手术组和心肌梗死植组各12只,结扎左冠状动脉前降支制备大鼠心肌梗死模型。心肌梗死后1周,经尾静脉注射脐血MSCs,4周后测定大鼠血流动力学。结果 42份脐血中单个核细胞原代培养,其中仅8份传代培养成功,而其中mesencult条件培养基有5份培养成功,明显高于FBS包被组(3份)和FBS未包被组(0份)。流式细胞仪检测第2代的脐血MSCs结果显示,P2代MSCs不表达或极弱表达CD 34,CD45造血细胞标志,稳定地高表达CD 29、CD 105间充质细胞相关的表面抗原标记。这与骨髓MSCs的表面抗原标志相一致。移植后4周,经血流动力学检测,与心梗组比较,MSCs移植组左室心功能明显改善(P<0.05)。结论将脐血单个核细胞以高密度(1×108cells/ml)接种在mesencult培养基中可以在体外成功培养出较纯化的脐血MSCs,其培养成功率较高。脐血MSCs的免疫表型符合间充质干细胞特征。脐血MSCs移植能促进心梗大鼠心功能恢复。

关键词：人脐带血间充质干细胞移植急性心肌梗死大鼠左室心功能 Acute Myocardial Infarction Heart Function Stem Cell Transplantation Mesenchymal umbilical Cord Blood 条件培养基脐血单个核细胞 MSCs移植流式细胞仪检测血流动力学检测细胞原代培养脐血间充质干细胞表面抗原心肌梗死模型体外培养体系

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NEWS AND NOTES

中华医学杂志(英文版) 1994年第5期 13+36-13+36页

CORD BLOOD HEMATOPOIETIC CELL BANK Human umbilical cord blood as a sufficient source of cells for hematopoietic cell transplantation, usually discarded,now has been used to restore the hematopoiesis of *** Medical University Hospital first developed the HLA-mismatched multi—cord blood transplantation and build up the first cord blood hematopoietic cell bank in the world. Studies have shown that the frequency of hematopoietic progenitors in cord blood equals to or even exceeds that of bone marrow and greatly surpasses that of adult blood. The cord blood can form CFU-F in culture besides CFU-GM and BFU-E. The frequency of CFU-GM in coculture of cord blood exceeds the sum of individual cultures. In has

关键词： hematopoietic NOTES umbilical Shandong restore CELL BANK besides 且一

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脐血CD34+细胞的分离与纯化

医学检验与临床 2007年第1期19卷 47-48,24页

作者：朱永宝王雪马京香山东省血液中心济南250014

脐血造血干细胞的分离是进行干细胞移植、体外扩增培养的关键。研究采用明胶自然沉降法、Ficoll分层法分离脐血MNC后,用MACS分离纯化CD34+细胞,计数并用流式细胞仪进行纯度分析。结果显示:每份脐血的采集量平均为95+52ml,3g/dl明胶自然... 详细信息

脐血造血干细胞的分离是进行干细胞移植、体外扩增培养的关键。研究采用明胶自然沉降法、Ficoll分层法分离脐血MNC后,用MACS分离纯化CD34+细胞,计数并用流式细胞仪进行纯度分析。结果显示:每份脐血的采集量平均为95+52ml,3g/dl明胶自然沉降法所获MNC密度平均为(5.76±0.67)×107/ml;CD34+细胞密度平均为(5.53±1.16)×105/ml,较Fi-coll分层法高(P<0.01)。因此,3g/dl明胶自然沉降法是一种较理想的分离MNC的方法,用明胶沉降法和MACS相结合是分离脐血CD34+细胞的理想方法。

关键词： umbilical cord blood CD34+ Separating method

Differential recognition of MHC class I molecules of xeno-/allo-endothelial cells by human NK cells

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Science China(Life Sciences) 2000年第2期43卷 176-182页

作者：冯志民张晓峰王宏芳丰美福 State Key Lab of Biomembrane and Membrane Biotechnology Institute of Zoology Chinese Academy of Sciences Beijing 100080 China

Using human umbilical vein endothelial cells (HUVEC) and porcine aortic endothelial cells (PAEC) as target cells, human peripheral blood NK cells (PBNK) and NK92 cells as effector cells, the differential cytotoxicities of NK cells to allo- and xeno-endothelial cells were studied. The influence of MHC class I molecules on the cytotoxicity of human NK cells was assayed using acid treatment, and blockades of MHC class I antigens, CD94 and KIR (NKB1). The results indicated that the killing of PAEC by the two kinds of NK cells is higher than that of HUVEC. After acid-treatment, the cytotoxicity of the two kinds of NK cells to PAEC and HUVEC is significantly enhanced, but the magnitude of the enhancement is different. The enhancement of NK killing to acid treated HUVEC is much greater than that to PAEC. Blockade of CD94 mAb did not alter the NK cytotoxicity, while blockade of NKB1 mAb enhanced the cytotoxicity of PBNK to HUVEC and PAEC by 95% and 29% respectively. The results above suggested that the differential recognition of MHC I molecules of xeno-endothelial cells by human NK cells could be the major reason for higher NK cytotoxicity to PAEC. KIR might be the primary molecule that transduced inhibitory signals when endothelial cells were injured by NK cells.

关键词： human umbilical vein endothelial cells (HUVEC) porcine aortic endothelial cells (PAEC) peripheral blood natural killer cells (PBNK) NK92 acid treatment MHC class I molecules cytotoxicity.

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Hemopoietic System

China Medical Abstracts(Internal Medicine) 2005年第3期22卷 176-185页

*** and erythrocyte function2005398 The effect of down regulating of P27 on theactivity of hematopoietic progenitor cells derivedfrom umbilical cord blood and bone marrow ex vivoLI Li(李莉),at *** Lab Med,Changzheng H... 详细信息

关键词： progenitor umbilical hematopoietic 李莉 regulating erythrocyte ATRA BMMNC isolation inhibited

Effect of Delayed versus Early Cord Clamping on Neonatal Outcomes and Iron Status at 4 Months

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Open Journal of Obstetrics and Gynecology 2019年第2期9卷 229-241页

作者： Musa O. Busarira Entesar Alasbaly Maryam Sulayman Mbark Department of Obstetrics and Gynecology Faculty of Medicine University of Benghazi Benghazi Libya Department of Obstetrics and Gynecology Ejdabia Hospital Ejdabia Libya

Background: The optimal timing of umbilical cord clamping has been debated in the scientific literature for over a century. Early clamping of the umbilical cord is widely practised as part of the management of labour. Early clamping of the cord was one of the first routine medical interventions in labour. Its place in modern births was guaranteed by its incorporation into the triad of interventions that make up the active management of the third stage of labour. Babies who have immediate cord clamping have also been shown to be more likely to be anaemic, have decreased fine motor skills, and have decreased social skills during their first few years of age. Delayed umbilical cord clamping appears to have clear benefit for term infants. Delayed umbilical cord clamping increases hemoglobin levels at birth and improves iron stores in the first few months of life, which improve the developmental outcomes. There is growing evidence that delayed cord clamping is beneficial and can improve the infant’s iron status for up to 6 months after birth. For the first few minutes after birth, there is still circulation from the placenta to the infant. Waiting to clamp the umbilical cord for 2 - 3 min, or until cord pulsations cease, allows a physiological transfer of placental blood to the infant (placental transfusion), the majority of which occurs within 3 min. This placental transfusion provides sufficient iron reserves for the first 6 - 8 months of life, preventing or delaying the development of iron deficiency until other interventions. WHO’s findings suggest that late cord clamping (one to three minutes after delivery or longer) is recommended for all births. Aim of the study: 1) To evaluate the benefit of delayed cord clamping on the infant’s health status. 2) To assess hemoglobin, iron and ferritin at birth and at 4 months of age in infants who underwent early cord clamping (at 30 seconds) as compared with infants who underwent delayed cord clamping (at 60 seconds). Methods:

关键词： Early umbilical Cord Clamping umbilical Cord Clamping Anaemia Hemoglobin Ferritin Polycythemia APGAR Score