28 - Session
Oral presentations Hemostasis, transfusion medicine, vascular, laboratory medicine, benign hematology
Nov. 22, 2023, 3:30 p.m. - 5:00 p.m., Shanghai 1-2
Modified Rotating Bed Reactor Compatible with an Industrializable and Scalable Manufacturing of Lab-Grown Platelets
M. Humbert1, A. Aliotta2, T. Badalli2, V. Tâche2, M. Després2, Y. Grand2, P. Jeandet2, V. Boand2, C. Rocca2, F. Khechana2, T. Braschler3, O. Naveiras1, E. Dahan2, L. Burnier2, Presenter: L. Burnier2 (1Lausanne, 2Saint-Sulpice, 3Geneva)
The increasing demand for platelet (PLT) transfusions requires a safe alternative to blood and PLT donation. In vitro production of platelets is currently explored, but achieving a commercially viable solution is hindered by the limitations of large-scale production and costs. This paper introduces a scalable solution that efficiently produces one platelet concentrate (PC) in under two hours, starting from mature megakaryocytes (MK).
Human CD34+ cells were seeded in a xeno-free medium containing thrombopoietin to differentiate into MK, and transferred into HemostOD’s modified rotating bed reactor – consisting of a pod containing microstructures – in a 200 mL bioreactor, for 2 hours at 900 RPM. Control PLTs were both from healthy donors (blood-PLT) or generated using a microfluidic chip (CHIP-PLT). PLT activation was done using ADP + PAR1 agonist peptide and analyzed by flow cytometry (FC). The pharmacokinetic study was done in immunodeficient clodronate-treated mice. Desialylation was measured using R. communis agglutinin. PLT (CD41+CD42b+) were counted by FC.
PLT generated in HemostOD’s bioreactor (HOD-PLT) were compared to CHIP-PLT and to blood-PLT. At resting state, 15.2+/-4.1% of CHIP-PLT were CD62P positive, while only 6.6+/-4.0% of HOD-PLTs were positive (mean+/-SD, n=3, P=0.057, 2-way ANOVA with Tukey multiple-comparisons test). Phosphatidylserine exposure was significantly reduced in HOD-PLTs (5.1+/-1.1%) compared to CHIP-PLTs (16.5+/-1.3%, P=0.01). Upon activation, PAC-1 binding and release of CD62P were increased in HOD-PLTs compared to CHIP-PLT (53.5+/-1.0% and 46.0+/-6.6% versus 24.0+/-4.1% and 32.0+/-1.1%, P<0.01). Sialylation (resting) and markers after activation of HOD-PLTs were similar to blood-PLT. After infusion in mice, survival of HOD-PLTs and blood-PLTs were also similar (half-life of 22+/-2 h and 17+/-2.0 h, respectively). Finally, 5,5*10^10 PLTs were produced using 4,5*10^9 MKs, reaching systematically a plateau between 90 and 120 minutes.
Functional platelets are generated in HemostOD’s bioreactor in 2 hours, sharing characteristics with blood-PLTs. HemostOD’s bioreactor in conjunction with HemostOD's proprietary human MK cell line thus has the potential to enable large-scale production of platelets for clinical use with an unlimited and stable source of megakaryocytes.