Platelets generated in vitro from proplatelet-displaying human megakaryocytes are functional

ES Choi, JL Nichol, MM Hokom, AC Hornkohl, P Hunt - 1995 - ashpublications.org
ES Choi, JL Nichol, MM Hokom, AC Hornkohl, P Hunt
1995ashpublications.org
An in vitro culture system demonstrating the transitions from megakaryocyte progenitors to
functional platelets is described. CD34-selected cells from normal human peripheral blood
are cultured under conditions that promote megakaryocyte formation. After 8 to 11 days,
enriched populations of mature megakaryocytes are replated under conditions that favor the
development of proplatelets. Proplatelets express the platelet-specific proteins,
glycoproteins Ib and IIb (GPIb and GPIIb), and fibrinogen and also contain microtubule coils …
Abstract
An in vitro culture system demonstrating the transitions from megakaryocyte progenitors to functional platelets is described. CD34- selected cells from normal human peripheral blood are cultured under conditions that promote megakaryocyte formation. After 8 to 11 days, enriched populations of mature megakaryocytes are replated under conditions that favor the development of proplatelets. Proplatelets express the platelet-specific proteins, glycoproteins Ib and IIb (GPIb and GPIIb), and fibrinogen and also contain microtubule coils equal in size to those found in plasma-derived platelets. In addition, proplatelets have ultrastructural features in common with plasma- derived platelets. Platelet-sized particles from the proplatelet culture supernatants are examined. Ultrastructurally, these particles are identical to plasma-derived platelets. Functionally, these culture- derived platelets aggregate in response to both thrombin and adenosine diphosphate (ADP) plus fibrinogen. This aggregation is specifically inhibited by the addition of a function-blocking anti-GPIIbIIIa antibody. Culture-derived platelets stimulated with agonists also express the activation-dependent antigens P-selectin and functional fibrinogen receptor. This is the first description of an in vitro culture system that sequentially demonstrates megakaryocyte growth, development, and platelet production.
ashpublications.org