title: Co-cultured hBMSCs and HUVECs on human bio-derived bone scaffolds provide support for the long-term ex vivo culture of HSC/HPCs.
abstract: In order to closely mimic a multi-cell state in hematopoietic stem/progenitor cells (HSC/HPCs) vascular niche, we co-cultured human bone marrow mesenchymal stem cells (hBMSCs) and human umbilical vein endothelial cells (HUVECs) without any cytokines as feeder cells and applied bio-derived bone from human femoral metaphyseal portion as scaffold to develop a new HSC/HPCs three-dimensional culture system (named 3D-Mix cultures). Scanning electron and fluorescent microscopy showed excellent biocompatibility of bio-derived bone to hBMSCs and HUVECs in vitro. Flow cytometry analysis and quantitative real-time polymerase chain reaction (qPCR) assay of p21 expression demonstrated that 3D-Mix could promote self-renewal and ex vivo expansion of HSCs/HPCs significantly higher than 3D-hMSC and 3D-HUVEC. Long-term culture initiating cell (LTC-IC) confirmed that 3D-Mix had the most powerful activity of maintaining multipotent differentiation of primitive cell subpopulation in HSCs. The nonobese diabetic/severe combined immunodeficiency (NOD/SCID) repopulating cell (SRC) assay demonstrated that 3D-Mix promoted the expansion of long-term primitive transplantable HSCs. qPCR of alkaline phosphatase (ALP) and osteocalcin (OC) demonstrated that HUVECs enhanced the early osteogenic differentiation of BMSCs. Western blot and qPCR revealed that HUVECs activated Wnt/β-catenin signaling in hBMSCs inducing Notch signal activation in HSCs. Our study indicated that interaction between hMSCs and HUVECs may have a critical role in to influent on HSCs/HPCs fate in vitro. These results demonstrated that the 3D-Mix have the ability to support the maintenance and proliferation of HSCs/HPCs in vitro.
authors.affiliation: Haematology Department, Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Huang
authors.affiliation: Haematology Department, Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Chenglong
authors.initials: C
authors.lastname: Li
authors.affiliation: Haematology Department, Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Biao
authors.initials: B
authors.lastname: Zhu
authors.affiliation: Center for Cell Transplantation (Seventh Unit of General Surgery Department), Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Hailian
authors.initials: H
authors.lastname: Wang
authors.affiliation: Center for Cell Transplantation (Seventh Unit of General Surgery Department), Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Xiangwei
authors.initials: X
authors.lastname: Luo
authors.affiliation: Center for Cell Transplantation (Seventh Unit of General Surgery Department), Institute of Organ Transplantation, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, 610072, People's Republic of China.
authors.forename: Lingling
authors.initials: L
authors.lastname: Wei
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002450:Cell Communication; D002454:Cell Differentiation; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D016513:Mice, SCID; D010012:Osteogenesis; D051880:Receptors, Notch / Q000378:metabolism; D015398:Signal Transduction; D054457:Tissue Scaffolds / Q000737:chemistry*; D060449:Wnt Signaling Pathway
chemical_list: D051880:Receptors, Notch
keywords: hematopoietic stem/progenitor cell (HSC/HPC); human bio-derived bone scaffold (hBDBS); human bone marrow mesenchymal stem cells (hBMSCs); human umbilical vein endothelial cells (HUVECs); three-dimensional (3D) culture system