title: Minimally manipulated whole human umbilical cord is a rich source of clinical-grade human mesenchymal stromal cells expanded in human platelet lysate.
abstract: OBJECTIVE
  Mesenchymal stromal cells (MSC) have recently been identified as a therapeutic option in several clinical conditions. Whereas bone marrow (BM) is considered the main source of MSC (BM-MSC), the invasive technique required for collection and the decline in allogeneic donations call for alternative sources. Human umbilical cord (UC) represents an easily available source of MSC (UC-MSC).
  
  
  METHODS
  Sections of full-term UC were transferred to cell culture flasks and cultured in 5% human platelet lysate (PL)-enriched medium. Neither enzymatic digestion nor blood vessel removal was performed. After 2 weeks, the adherent cells were harvested (P1), replated at low density and expanded for two consecutive rounds (P2 and P3).
  
  
  RESULTS
  We isolated and expanded MSC from 9/9 UC. UC-MSC expanded with a mean fold increase (FI) of 42 735 ± 16 195 from P1 to P3 in a mean of 29 ± 2 days. By processing the entire cord unit, we theoretically could have reached a median of 9.5 × 10(10) cells (ranging from 1.0 × 10(10) to 29.0 × 10(10)). UC-MSC expressed standard surface markers; they contained more colony-forming unit (CFU)-fibroblast (F) and seemed less committed towards osteogenic, chondrogenic and adipogenic lineages than BM-MSC. They showed immunosuppressive properties both in vitro and in an in vivo chronic Graft versus Host disease (cGvHD) mouse model. Both array-Comparative Genomic Hybridization (CGH) analysis and karyotyping revealed no chromosome alterations at the end of the expansion. Animal studies revealed no tumorigenicity in vivo.
  
  
  CONCLUSIONS
  UC constitute a convenient and very rich source of MSC for the production of third-party 'clinical doses' of cells under good manufacturing practice (GMP) conditions.
authors.affiliation: Laboratory of Cell Therapy G. Lanzani, USC Hematology, Ospedali Riuniti di Bergamo, Bergamo, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Capelli
authors.forename: Elisa
authors.initials: E
authors.lastname: Gotti
authors.forename: Marina
authors.initials: M
authors.lastname: Morigi
authors.forename: Cinzia
authors.initials: C
authors.lastname: Rota
authors.forename: Ling
authors.initials: L
authors.lastname: Weng
authors.forename: Francesco
authors.initials: F
authors.lastname: Dazzi
authors.forename: Orietta
authors.initials: O
authors.lastname: Spinelli
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cazzaniga
authors.forename: Rosangela
authors.initials: R
authors.lastname: Trezzi
authors.forename: Andrea
authors.initials: A
authors.lastname: Gianatti
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rambaldi
authors.forename: Josee
authors.initials: J
authors.lastname: Golay
authors.forename: Martino
authors.initials: M
authors.lastname: Introna
mesh_terms: D050156:Adipogenesis; D000818:Animals; D001792:Blood Platelets / Q000166:cytology; D015197:Carcinogenicity Tests; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D055028:Comparative Genomic Hybridization; D003470:Culture Media; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000166:cytology; D006086:Graft vs Host Disease / Q000276:immunology; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D013234:Stem Cells; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D003470:Culture Media; D007166:Immunosuppressive Agents