title: Ex vivo differentiation of cord blood stem cells into megakaryocytes and platelets.
abstract: Megakaryocytes (MK) are hematopoietic cells present in the bone marrow that are responsible for the production and release of platelets in the circulation. Given their very low frequency (<1%), human MK often need to be derived in culture to study their development or to generate sufficient material for biological studies. This chapter describes a simplified 14-day culture protocol that efficiently leads to the production of MK and platelets from cord blood enriched progenitor cells. A serum-free medium is suggested for the growth of the CB cells together with an optimized cytokine cocktail developed specifically for MK differentiation, expansion, and maturation. Methodologies for flow cytometry analysis, MK and platelets estimation, and MK progenitor assay are also presented.
authors.affiliation: Département de Recherche et Développement, Héma-Québec, Université Laval, Québec City, QC, Canada. nicolas.pineault@hema-quebec.qc.ca
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pineault
authors.forename: Amélie
authors.initials: A
authors.lastname: Robert
authors.forename: Valérie
authors.initials: V
authors.lastname: Cortin
authors.forename: Lucie
authors.initials: L
authors.lastname: Boyer
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D003114:Colony-Forming Units Assay / Q000379:methods*; D015925:Cryopreservation; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D011003:Ploidies; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D018952:Antigens, CD34