title: Cell viability evaluation of transdifferentiated endothelial-like cells by quantitative electron-probe X-ray microanalysis for tissue engineering.
abstract: Development of an efficient vascular substitute by tissue engineering is strongly dependent on endothelial cell viability. The aim of this study was to evaluate cell viability of transdifferentiated endothelial-like cells (Tr-ELC) by using for the first time electron probe X-ray microanalysis (EPXMA), not only to accurately analyze cell viability by quantifying the intracellular ionic concentrations, but also to establish their possible use in vascular tissue engineering protocols. Human umbilical cord Wharton's jelly stem cells (HWJSC) and endothelial cells from the human umbilical vein (HUVEC) were isolated and cultured. Transdifferentiation from HWJSC to the endothelial phenotype was induced. EPXMA was carried out to analyze HUVEC, HWJSC and Tr-ELC cells by using a scanning electron microscope equipped with an EDAX DX-4 microanalytical system and a solid-state backscattered electron detector. To determine total ion content, the peak-to-local-background (P/B) ratio method was used with reference to standards composed of dextran containing known amounts of inorganic salts. Our results revealed a high K/Na ratio in Tr-ELC (9.41), in association with the maintenance of the intracellular levels of chlorine, phosphorous and magnesium and an increase of calcium (p=0.031) and sulfur (p=0.022) as compared to HWJSC. Calcium levels were similar for HUVEC and Tr-ELC. These results ensure that transdifferentiated cells are highly viable and resemble the phenotypic and microanalytical profile of endothelial cells. Tr-ELC induced from HWJSC may fulfill the requirements for use in tissue engineering protocols applied to the vascular system at the viability and microanalytical levels.
authors.affiliation: Faculty of Health Sciences and Tondela-Viseu Hospital, University of Beira Interior, Covilhã, Viseu, Portugal.
authors.forename: Manuel
authors.initials: M
authors.lastname: Vico
authors.affiliation: Division of Vascular Surgery, Virgen de la Victoria University Hospital, Málaga, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Rodríguez-Morata
authors.affiliation: Department of Histology (Tissue Engineering Group), University of Granada, Granada, Spain.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Garzón
authors.affiliation: Department of Histology (Tissue Engineering Group), University of Granada, Granada, Spain.
authors.forename: Fernando
authors.initials: F
authors.lastname: Campos
authors.affiliation: Department of Histology (Tissue Engineering Group), University of Granada, Granada, Spain.
authors.forename: Boris
authors.initials: B
authors.lastname: Jaimes-Parra
authors.affiliation: Department of Medical Specialities, Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), University of Alcalá, Alcalá de Henares, Spain.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pérez-Köhler
authors.affiliation: Department of Medical Specialities, Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), University of Alcalá, Alcalá de Henares, Spain.
authors.forename: Julia
authors.initials: J
authors.lastname: Buján
authors.affiliation: Department of Histology (Tissue Engineering Group), University of Granada, Granada, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Alaminos
authors.affiliation: Department of Histology (Tissue Engineering Group), University of Granada, Granada, Spain. mcsanchez@ugr.es.
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Sánchez-Quevedo
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation; D002470:Cell Survival; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D004577:Electron Probe Microanalysis; D066026:Endothelial Progenitor Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D010641:Phenotype; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D015415:Biomarkers