BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an ...BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.展开更多
The effects of mifepristone with misoprostol on the expression of the integrin β 3 and intercellular adhesion molecule 1 (ICAM 1) in decidua and chorionic villi tissues in early pregnancy in 10 cases were investigate...The effects of mifepristone with misoprostol on the expression of the integrin β 3 and intercellular adhesion molecule 1 (ICAM 1) in decidua and chorionic villi tissues in early pregnancy in 10 cases were investigated by immuno flow cytometry (the experiment group). At the same time, the other 10 cases induced by mechanical vacuum aspiration were collected as the control. The results showed that, the positive rate of integrin β 3 and ICAM 1 in decidua of the experiment group were 19.1±5.01% and 20.61 ±6.51%; while those in chorionic villi were 21.32±4.38% and 20.29± 6.49%, which were significantly lower than those in the control group. These results suggested that integrin β 3 and ICAM 1 may take part in the maintenance of early pregnancy. The mechanism of mifepristone induced abortion may be mediated by the down regulation of the integrin β 3 and ICAM 1 expression in decidua and chorionic villi.展开更多
Objective:This study aims to explore its role in embryo damage and the relationship between them by testing the expression of PCNA,Caspase-3,IL-6 and Survivin protein in the chorionic villi and decidual tissue of pati...Objective:This study aims to explore its role in embryo damage and the relationship between them by testing the expression of PCNA,Caspase-3,IL-6 and Survivin protein in the chorionic villi and decidual tissue of patients with unexplained early embryo damage and normal early pregnancy at the same time voluntarily requested uterine aspiration abortion in order to clarify the pathogenesis of early embryo damage from the cellular and molecular perspectives,and provide theoretical basis for the diagnosis and treatment of embryo damage.Methods:From July 2018 to July 2019,30 patients with unexplained embryo damage were selected as embryo damage group,thirty normal early pregnancy patients with aspiration abortion were selected as the normal early pregnancy group.The decidual tissue and chorionic villi of the two groups were collected to observe the structural and pathological changes.Immunohistochemical method was used to detect the distribution of PCNA,Caspase-3,IL-6 and Survivin in the chorionic villi and decidual tissue,as well as the expression changes of the above four proteins.Results:Compared with the normal early pregnancy group,the chorionic villi in the early embryo damage group were obviously dysplasia,degenerative changes,structural disorders,homogeneous destruction,obvious reduction or even disappearance of trophoblast cells,inflammatory cell infiltration,more neutrophils and lymphocytes,interstitial edema and cell atrophy,accompanied by hemorrhage;The decidual tissue of the intima was not good,most of the cells were spindle-shaped,the structure was disordered,the stroma has edema,inflammatory cells can be seen,and hemorrhage existed.The results of immunohistochemistry showed that the expression of PCNA protein in chorionic villi and decidual tissue of early embryo damage group decreased significantly(P<0.05),while the expression of Caspase-3 protein increased significantly(P<0.05).The expression of IL-6 protein decreased significantly(P<0.05),and the expression of Survivin protein decreased significantly(P<0.05).Conclusion:During early pregnancy,due to the down-regulation of the expression of proliferating cell nuclear antigen PCNA and apoptosis inhibitor protein Survivin in chorionic and decidual tissues,the balance between proliferation and apoptosis is broken,which has an increase in Caspase-3 expression and a decrease in IL-6 expression.It may be that the balance of Th1/Th2 in chorionic and decidual tissues inclines to Th1,which leads to excessive apoptosis of chorionic and decidual tissues and the cessation of early embryo damage.展开更多
目的：评价改良荧光原位杂交（fluorescent in situ hybridization,FISH）技术在产前诊断中的应用。方法：用改良FISH技术检测119例孕16-24周孕妇的羊水间期细胞及10例孕25-32周胎儿脐血间期细胞,5例孕9-12周绒毛间期细胞,每例均行常规...目的：评价改良荧光原位杂交（fluorescent in situ hybridization,FISH）技术在产前诊断中的应用。方法：用改良FISH技术检测119例孕16-24周孕妇的羊水间期细胞及10例孕25-32周胎儿脐血间期细胞,5例孕9-12周绒毛间期细胞,每例均行常规染色体核型分析。结果：应用改良FISH法,所有样本均在6h内获得检测结果,除2例羊水培养失败外,其余样本均在3周内获得细胞遗传学诊断。两种方法均检出特氏综合征、18-三体综合征、21-三体综合征各1例,另5例常规染色体核型分析异常,因超出检测范围,FISH法未能检出,所有样本的两种方法检测结果均一致。结论：经改良后的FISH技术缩短了诊断时间,缓解了孕妇及家属的焦虑心情,且可用于多种不同样本的检测,因其高效、省时、取材多样等优点在产前诊断具有重要的临床价值。展开更多
基金the Shriners Hospital for Children Postdoctoral Research Fellowship award,No.84704-NCA-19UC Davis School of Medicine Dean’s Fellowship award and funding from the NIH,No.5R01NS100761-02 and No.R03HD091601-01+2 种基金the California Institute of Regenerative Medicine,No.PC1-08103 and No.CLIN1-11404Shriners Hospitals for Children,No.85120-NCA-16,No.85119-NCA-18,No.85108-NCA-19 and No.87200-NCA-19March of Dimes Foundation,No.5FY1682
文摘BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.
文摘The effects of mifepristone with misoprostol on the expression of the integrin β 3 and intercellular adhesion molecule 1 (ICAM 1) in decidua and chorionic villi tissues in early pregnancy in 10 cases were investigated by immuno flow cytometry (the experiment group). At the same time, the other 10 cases induced by mechanical vacuum aspiration were collected as the control. The results showed that, the positive rate of integrin β 3 and ICAM 1 in decidua of the experiment group were 19.1±5.01% and 20.61 ±6.51%; while those in chorionic villi were 21.32±4.38% and 20.29± 6.49%, which were significantly lower than those in the control group. These results suggested that integrin β 3 and ICAM 1 may take part in the maintenance of early pregnancy. The mechanism of mifepristone induced abortion may be mediated by the down regulation of the integrin β 3 and ICAM 1 expression in decidua and chorionic villi.
文摘Objective:This study aims to explore its role in embryo damage and the relationship between them by testing the expression of PCNA,Caspase-3,IL-6 and Survivin protein in the chorionic villi and decidual tissue of patients with unexplained early embryo damage and normal early pregnancy at the same time voluntarily requested uterine aspiration abortion in order to clarify the pathogenesis of early embryo damage from the cellular and molecular perspectives,and provide theoretical basis for the diagnosis and treatment of embryo damage.Methods:From July 2018 to July 2019,30 patients with unexplained embryo damage were selected as embryo damage group,thirty normal early pregnancy patients with aspiration abortion were selected as the normal early pregnancy group.The decidual tissue and chorionic villi of the two groups were collected to observe the structural and pathological changes.Immunohistochemical method was used to detect the distribution of PCNA,Caspase-3,IL-6 and Survivin in the chorionic villi and decidual tissue,as well as the expression changes of the above four proteins.Results:Compared with the normal early pregnancy group,the chorionic villi in the early embryo damage group were obviously dysplasia,degenerative changes,structural disorders,homogeneous destruction,obvious reduction or even disappearance of trophoblast cells,inflammatory cell infiltration,more neutrophils and lymphocytes,interstitial edema and cell atrophy,accompanied by hemorrhage;The decidual tissue of the intima was not good,most of the cells were spindle-shaped,the structure was disordered,the stroma has edema,inflammatory cells can be seen,and hemorrhage existed.The results of immunohistochemistry showed that the expression of PCNA protein in chorionic villi and decidual tissue of early embryo damage group decreased significantly(P<0.05),while the expression of Caspase-3 protein increased significantly(P<0.05).The expression of IL-6 protein decreased significantly(P<0.05),and the expression of Survivin protein decreased significantly(P<0.05).Conclusion:During early pregnancy,due to the down-regulation of the expression of proliferating cell nuclear antigen PCNA and apoptosis inhibitor protein Survivin in chorionic and decidual tissues,the balance between proliferation and apoptosis is broken,which has an increase in Caspase-3 expression and a decrease in IL-6 expression.It may be that the balance of Th1/Th2 in chorionic and decidual tissues inclines to Th1,which leads to excessive apoptosis of chorionic and decidual tissues and the cessation of early embryo damage.
文摘目的：评价改良荧光原位杂交（fluorescent in situ hybridization,FISH）技术在产前诊断中的应用。方法：用改良FISH技术检测119例孕16-24周孕妇的羊水间期细胞及10例孕25-32周胎儿脐血间期细胞,5例孕9-12周绒毛间期细胞,每例均行常规染色体核型分析。结果：应用改良FISH法,所有样本均在6h内获得检测结果,除2例羊水培养失败外,其余样本均在3周内获得细胞遗传学诊断。两种方法均检出特氏综合征、18-三体综合征、21-三体综合征各1例,另5例常规染色体核型分析异常,因超出检测范围,FISH法未能检出,所有样本的两种方法检测结果均一致。结论：经改良后的FISH技术缩短了诊断时间,缓解了孕妇及家属的焦虑心情,且可用于多种不同样本的检测,因其高效、省时、取材多样等优点在产前诊断具有重要的临床价值。
