Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration  

作　　者：Kewa Gao Siqi He Priyadarsini Kumar Diana Farmer Jianda Zhou Aijun Wang 

机构地区：[1]Department of Burns and Plastic Surgery,The Third Xiangya Hospital of Central South University,Changsha 410013,Hunan Province,China [2]Surgical Bioengineering Laboratory,Department of Surgery,University of California Davis,Sacramento,CA 95817,United States [3]Institute for Pediatric Regenerative Medicine,Shriners Hospitals for Children,Sacramento,CA 95817,United States [4]Department of Biomedical Engineering,University of California Davis,Davis,CA 95817,United States

出　　处：《World Journal of Stem Cells》2020年第2期123-138,共16页世界干细胞杂志（英文版）（电子版）

基　　金：the Shriners Hospital for Children Postdoctoral Research Fellowship award,No.84704-NCA-19;UC Davis School of Medicine Dean’s Fellowship award and funding from the NIH,No.5R01NS100761-02 and No.R03HD091601-01;the California Institute of Regenerative Medicine,No.PC1-08103 and No.CLIN1-11404;Shriners Hospitals for Children,No.85120-NCA-16,No.85119-NCA-18,No.85108-NCA-19 and No.87200-NCA-19;March of Dimes Foundation,No.5FY1682

摘　　要：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 ECFCsBACKGROUND 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/td Tomato-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 × 10~7 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,simila

关 键 词：PLACENTA Endothelial colony forming cells Chorionic villi Angiogenesis Tissue engineering 

分 类 号：R714.5[医药卫生—妇产科学]