title: Absence of micronucleus formation in CHO-K1 cells cultivated in platelet lysate enriched medium.
abstract: Human platelet lysate (PL) represents an effective substitute of fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) cultivation. Compared to FBS, PL favors MSC proliferation significantly shortening the population doubling time and avoiding the risks related to the use of animal derivatives. Growth factors contained in the platelets are released upon platelet disruption following freezing/thawing cycles or as we have recently described by using ultrasound. We have investigated whether the increased cell proliferation achieved by using PL could induce mitotic stress and whether the potential formation of free radicals during PL production by ultrasound could cause chromosomal instability in mammalian cells. We have applied an image analysis assisted high content screening (HCS) in vitro micronucleus assay in the Chinese Hamster Ovarian K1 (CHO-K1) rodent mammalian cell line. PL was produced by sonication; for the micronucleus assay, CHO-K1 cells were exposed to increasing concentrations of PL. Cytokinesis was blocked by cytochalasin B, nuclei were stained with bisbenzimide and images were acquired and analyzed automatically using an HCS system, both with a 20× and a 10× objective. Our results suggest that growth stimulus induced by the use of PL did not significantly increase micronucleus formation in CHO-K1 cells compared to negative control. Micronucleus testing in conjunction with HCS could represent a valid tool to evaluate the safety of ancillary materials used in the production of cell-based medicinal products.
authors.affiliation: Hematology Project Foundation Research Laboratories, Contrà S. Francesco 41, Vicenza, Italy; Advanced Cellular Therapy Laboratory, Department of Cellular Therapy and Hematology, San Bortolo Hospital, Via Rodolfi 37, Vicenza, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Bernardi
authors.affiliation: High Throughput Screening Core Facility, CIBIO (Centre for Integrative Biology) - University of Trento, via delle Regole 101, Mattarello (TN), Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Adami
authors.affiliation: Hematology Project Foundation Research Laboratories, Contrà S. Francesco 41, Vicenza, Italy; Advanced Cellular Therapy Laboratory, Department of Cellular Therapy and Hematology, San Bortolo Hospital, Via Rodolfi 37, Vicenza, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Albiero
authors.affiliation: Hematology Project Foundation Research Laboratories, Contrà S. Francesco 41, Vicenza, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Madeo
authors.affiliation: Hematology Project Foundation Research Laboratories, Contrà S. Francesco 41, Vicenza, Italy; Advanced Cellular Therapy Laboratory, Department of Cellular Therapy and Hematology, San Bortolo Hospital, Via Rodolfi 37, Vicenza, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Rodeghiero
authors.affiliation: Advanced Cellular Therapy Laboratory, Department of Cellular Therapy and Hematology, San Bortolo Hospital, Via Rodolfi 37, Vicenza, Italy. Electronic address: astori@hemato.ven.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Astori
mesh_terms: D000818:Animals; D001792:Blood Platelets; D016466:CHO Cells; D018929:Cell Culture Techniques; D002457:Cell Extracts / Q000494:pharmacology*; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D006801:Humans; D007091:Image Processing, Computer-Assisted; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects* / Q000706:statistics & numerical data; D015162:Micronucleus Tests; D053657:Platelet-Rich Plasma; D013010:Sonication
chemical_list: D002457:Cell Extracts; D016895:Culture Media, Serum-Free
keywords: CHO-K1; High content screening; Mesenchymal stromal cells; Micronucleus; Platelet lysate; Platelet sonication