title: Evaluation of a new standardized enzymatic isolation protocol for human umbilical cord-derived stem cells.
abstract: The umbilical cord (UC) represents an important source of mesenchymal stem cells (MSC). These human UC-derived MSC (UC-MSC) have already been isolated using a protocol based on the migratory and plastic adhesive properties of MSC (UC-MSC-Mig). The UC-MSC-Mig isolation method, however, is difficult to standardize. Therefore, we developed an enzymatic isolation protocol (UC-MSC-Enz) to overcome the above mentioned disadvantages. First, we investigated the UC-MSC-Enz for their MSC properties. We found that UC-MSC-Enz express the MSC markers CD73, CD90 and CD105 and are able to differentiate into osteoblasts, adipocytes and chondroblasts fulfilling the MSC criteria of the International Society for Cellular Therapy. Previously we found that UC-MSC-Mig are unique among MSCs due to their significant expression of several hepatic (progenitor) markers. Therefore, we also investigated the expression of hepatic transcription factors and other hepatic markers in UC-MSC-Enz at both the mRNA and protein level. We found that the expression of hepatic transcription factors (GATA4, GATA6, SOX9 and SOX17) and hepatic markers (AFP, DPP4, CX43, DKK1, DSG2, KRT18 and KRT19) in UC-MSC-Enz was not significantly different from those of UC-MSC-Mig. Consequently, this optimized enzyme-based method represents a fast, robust and standardized way to isolate UC-MSC for a broad range of applications.
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium. Electronic address: karobuyl@vub.ac.be.
authors.forename: Karolien
authors.initials: K
authors.lastname: Buyl
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium.
authors.forename: Tamara
authors.initials: T
authors.lastname: Vanhaecke
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium.
authors.forename: Terry
authors.initials: T
authors.lastname: Desmae
authors.affiliation: Laboratory of Clinical Cell Therapy, Bordet Institut, Université Libre de Bruxelles, 808 Route de Lennik, B-1070 Brussels, Belgium.
authors.forename: Laurence
authors.initials: L
authors.lastname: Lagneaux
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium.
authors.forename: Vera
authors.initials: V
authors.lastname: Rogiers
authors.affiliation: Laboratory of Clinical Cell Therapy, Bordet Institut, Université Libre de Bruxelles, 808 Route de Lennik, B-1070 Brussels, Belgium.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Najar
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090 Brussels, Belgium.
authors.forename: Joery
authors.initials: J
authors.lastname: De Kock
mesh_terms: D002454:Cell Differentiation; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism; D051301:Desmoglein 2 / Q000235:genetics / Q000378:metabolism; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D053538:Keratin-18 / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D020784:Matrix Metalloproteinase 8 / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology*; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018031:Connexin 43; C501126:DKK1 protein, human; C495314:DSG2 protein, human; D051301:Desmoglein 2; D036341:Intercellular Signaling Peptides and Proteins; C488421:KRT18 protein, human; D053538:Keratin-18; D014157:Transcription Factors; D000509:alpha-Fetoproteins; C042807:DPP4 protein, human; D018819:Dipeptidyl Peptidase 4; D020784:Matrix Metalloproteinase 8
keywords: Hepatocyte; Isolation; Liver development; Mesenchymal stem cell; Umbilical cord