Uniform Impact on Individual Megakaryocytes is Essential for Efficient in Vitro Platelet Production

Overview

Journal Sci Rep

Date 2025 Jan 13

PMID 39805910

Authors

Andrei K Garzon Dasgupta

Anais Pongerard

Lea Mallo

Anita Eckly

Francois Lanza

Olivier Boiron

Yannick Knapp

Catherine Strassel

Affiliations

Soon will be listed here.

Abstract

Different approaches are being developed to efficiently produce in vitro platelets from cultured megakaryocytes to meet the constant demand of platelet transfusion and serve for research purposes. Recent works have shown that turbulence and periodic stress can significantly enhance platelet yield. Here we have developed and characterized a platelet production device that takes in account these properties. This device is based on the Taylor-Couette reactor in which a suspension is confined and sheared between two concentric cylinders. We have demonstrated that such a system allows obtaining high number of in vitro platelets per megakaryocyte with native-like morphology and functional properties. Using the combination of in silico and in vitro techniques, we claimed that overall turbulent conditions are not sufficient for efficient platelet release, and highlighted the importance of the uniform impact of flow on each megakaryocyte, a property that must be taken into account along with general flow characteristics when designing platelet release bioreactors. In addition, we have demonstrated that our system can be scaled up to large volumes without loss of efficiency, a significant advantage for the industrialization of platelet culture. In conclusion, we have developed a platelet production device with a predictable and highly precise effect on each megakaryocyte.

References

Bacher C, Bender M, Gekle S . Flow-accelerated platelet biogenesis is due to an elasto-hydrodynamic instability. Proc Natl Acad Sci U S A. 2020; 117(32):18969-18976. PMC: 7431004. DOI: 10.1073/pnas.2002985117. View

Strassel C, Lanza F, Gachet C . [Cultured platelets]. Bull Acad Natl Med. 2020; 204(9):971-980. PMC: 7556249. DOI: 10.1016/j.banm.2020.10.002. View

Li H, Liu Z, Lu L, Buffet P, Karniadakis G . How the spleen reshapes and retains young and old red blood cells: A computational investigation. PLoS Comput Biol. 2021; 17(11):e1009516. PMC: 8584971. DOI: 10.1371/journal.pcbi.1009516. View

Qi Y, Tan S, Corbitt N, Urbanik C, Salibindla A, Ni R . Fragmentation in turbulence by small eddies. Nat Commun. 2022; 13(1):469. PMC: 8789894. DOI: 10.1038/s41467-022-28092-3. View

Moreau T, Evans A, Vasquez L, Tijssen M, Yan Y, Trotter M . Large-scale production of megakaryocytes from human pluripotent stem cells by chemically defined forward programming. Nat Commun. 2016; 7:11208. PMC: 4829662. DOI: 10.1038/ncomms11208. View

Chen S, Sugimoto N, Eto K . Ex vivo manufacturing of platelets: beyond the first-in-human clinical trial using autologous iPSC-platelets. Int J Hematol. 2022; 117(3):349-355. PMC: 9792917. DOI: 10.1007/s12185-022-03512-8. View

Pongerard A, Mallo L, Gachet C, de la Salle H, Lanza F, Strassel C . Leukodepletion Filters-Derived CD34+ Cells As a Cell Source to Study Megakaryocyte Differentiation and Platelet Formation. J Vis Exp. 2021; (171). DOI: 10.3791/62499. View

Kim S, Ong P, Yalcin O, Intaglietta M, Johnson P . The cell-free layer in microvascular blood flow. Biorheology. 2009; 46(3):181-9. DOI: 10.3233/BIR-2009-0530. View

Lefrancais E, Ortiz-Munoz G, Caudrillier A, Mallavia B, Liu F, Sayah D . The lung is a site of platelet biogenesis and a reservoir for haematopoietic progenitors. Nature. 2017; 544(7648):105-109. PMC: 5663284. DOI: 10.1038/nature21706. View

10.

Pongerard A, Mallo L, Do Sacramento V, Boiron O, Eckly A, Gachet C . Development of an efficient, ready to use, blood platelet-release device based on two new flow regime parameters: The periodic hydrodynamic loading and the shear stress accumulation. N Biotechnol. 2023; 77:68-79. DOI: 10.1016/j.nbt.2023.07.002. View

11.

Broos K, Feys H, De Meyer S, Vanhoorelbeke K, Deckmyn H . Platelets at work in primary hemostasis. Blood Rev. 2011; 25(4):155-67. DOI: 10.1016/j.blre.2011.03.002. View

12.

Palma-Barqueros V, Revilla N, Sanchez A, Zamora Canovas A, Rodriguez-Alen A, Marin-Quilez A . Inherited Platelet Disorders: An Updated Overview. Int J Mol Sci. 2021; 22(9). PMC: 8123627. DOI: 10.3390/ijms22094521. View

13.

Javadi E, Deng Y, Karniadakis G, Jamali S . In silico biophysics and hemorheology of blood hyperviscosity syndrome. Biophys J. 2021; 120(13):2723-2733. PMC: 8390893. DOI: 10.1016/j.bpj.2021.05.013. View

14.

Hirata S, Murata T, Suzuki D, Nakamura S, Jono-Ohnishi R, Hirose H . Selective Inhibition of ADAM17 Efficiently Mediates Glycoprotein Ibα Retention During Ex Vivo Generation of Human Induced Pluripotent Stem Cell-Derived Platelets. Stem Cells Transl Med. 2017; 6(3):720-730. PMC: 5442763. DOI: 10.5966/sctm.2016-0104. View

15.

Do Sacramento V, Mallo L, Freund M, Eckly A, Hechler B, Mangin P . Functional properties of human platelets derived in vitro from CD34 cells. Sci Rep. 2020; 10(1):914. PMC: 6976668. DOI: 10.1038/s41598-020-57754-9. View

16.

Wool G, Miller J . The Impact of COVID-19 Disease on Platelets and Coagulation. Pathobiology. 2020; 88(1):15-27. PMC: 7649697. DOI: 10.1159/000512007. View

17.

Ma R, Bi Y, Kou J, Zhou J, Shi J . Enhanced procoagulant activity of platelets after chemotherapy in non-small cell lung cancer. Cancer Biol Ther. 2017; 18(8):627-634. PMC: 5653186. DOI: 10.1080/15384047.2017.1345387. View

18.

Ito Y, Nakamura S, Sugimoto N, Shigemori T, Kato Y, Ohno M . Turbulence Activates Platelet Biogenesis to Enable Clinical Scale Ex Vivo Production. Cell. 2018; 174(3):636-648.e18. DOI: 10.1016/j.cell.2018.06.011. View

19.

Trowbridge E, Martin J, Slater D, Kishk Y, Warren C, Harley P . The origin of platelet count and volume. Clin Phys Physiol Meas. 1984; 5(3):145-70. DOI: 10.1088/0143-0815/5/3/007. View

20.

Mookerjee S, Foster H, Waller A, Ghevaert C . In vitro-derived platelets: the challenges we will have to face to assess quality and safety. Platelets. 2020; 31(6):724-730. DOI: 10.1080/09537104.2020.1769051. View