title: Dissection of the cord blood stromal component reveals predictive parameters for culture outcome.
abstract: In regenerative medicine, human cord blood-derived multipotent mesenchymal stromal cells (CBMSCs) stand out for their biological peculiarities demonstrated in in vitro and in vivo preclinical studies. Here, we present our 9-year experience for the consistent isolation of CBMSCs. Although nearly one CB unit out of two retains the potential to give rise to MSC colonies, only 46% of them can be cultured till low passages (P≥4), but one-fourth of those reaches even higher passages (P≥8). Subsequent characterization for morphological, clonal, differentiation, and proliferation properties revealed two divergent CBMSC behaviors. In particular, a cumulative population doublings cut-off (CPD=15) was identified that undoubtedly distinguishes two growth curves, and different degrees of commitment toward osteogenesis were observed. These data clearly show the existence of at least two distinct CBMSC subsets: one mainly short-living and less proliferative (SL-CBMSCs), the other long-living, with higher growth rate, and, very importantly, with significantly (P≤0.01) longer telomere (LL-CBMSCs). Moreover, significant differences in the immunoprofile before seeding were found among CB units giving rise to LL-CBMSCs or SL-CBMSCs or showing no colony formation. Finally, all the aforementioned results provided a peculiar and useful set of parameters potentially predictive for CBMSC culture outcome.
authors.affiliation: 1 Cell Factory, Unit of Cell Therapy and Cryobiology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico , Milano, Italy .
authors.forename: Mario
authors.initials: M
authors.lastname: Barilani
authors.forename: Cristiana
authors.initials: C
authors.lastname: Lavazza
authors.forename: Mariele
authors.initials: M
authors.lastname: Viganò
authors.forename: Tiziana
authors.initials: T
authors.lastname: Montemurro
authors.forename: Valentina
authors.initials: V
authors.lastname: Boldrin
authors.forename: Valentina
authors.initials: V
authors.lastname: Parazzi
authors.forename: Elisa
authors.initials: E
authors.lastname: Montelatici
authors.forename: Mariacristina
authors.initials: M
authors.lastname: Crosti
authors.forename: Monica
authors.initials: M
authors.lastname: Moro
authors.forename: Rosaria
authors.initials: R
authors.lastname: Giordano
authors.forename: Lorenza
authors.initials: L
authors.lastname: Lazzari
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010012:Osteogenesis