OC 39.3 - Platelet Biogenesis from Mouse Megakaryocytes at Variable Shear Stress Levels by Using Microfluidic Systems

Monday, June 26, 2023

11:15 – 11:30 ET

Room: 515

Session Available As: On-Demand

Co-Author(s)

Semra Z. Birol, PhD

Postdoctoral Research Associate
School of Physiology, Pharmacology and Neuroscience/University of Bristol
Bristol, England, United Kingdom

Presenting Author(s)

Jordan Vautrinot, Bsc

phd student
University of bristol
Malvern, England, United Kingdom

Co-Author(s)

XZ

Xiaojuan Zhao

Senior Research Associate
School of Physiology, Pharmacology and Neuroscience/University of Bristol
Bristol, England, United Kingdom
MP

Muhammed E. Polat

PhD Student
Faculty of Mechanical Engineering/Istanbul Technical University
Istanbul, Istanbul, Turkey
SC

Sertac Cadirci

Associate Professor
Faculty of Mechanical Engineering/Istanbul Technical University
Istanbul, Istanbul, Turkey
LT

Levent Trabzon

Professor of Faculty of Mechanical Engineering
Faculty of Mechanical Engineering/Istanbul Technical University
Istanbul, Istanbul, Turkey
AP

Alastair W. Poole

Principal Investigator
School of Physiology, Pharmacology and Neuroscience/University of Bristol
Bristol, England, United Kingdom

Background: An ageing population, haematological malignancies, and a critical shortage of registered donors have contributed to considerable strain and the requirement for readily available platelet stocks. Therefore, donor-independent platelet biogenesis circumvents complications associated with traditional allogeneic platelet donation and transfusions such as a short shelf-life and the risk of bloodborne infections.

Aims: Two microfluidic systems have been designed for potential use in the generation of large quantities of platelets in a consistent and cost-effective manner. The initial system is primarily based on the structure of the pulmonary microvascular network, as it has been reported that platelet biogenesis can occur in the pulmonary capillary network. The second microfluidic system mimics better the physiological shear stress and pressure of the pulmonary capillary system.

Methods: We fabricated the microfluidic systems from Polydimethylsiloxane and glass through soft lithography, simulating the performance through Computational Fluid Dynamics. Mouse megakaryocytes, differentiated from mouse bone marrow, were isolated, stained with anti-CD41, and then passaged, at different flow rates through each system a total of six times (inspired by preliminary experiments). The number of generated platelets was quantified using Flow Cytometry and the resultant platelets were examined by Transmission Electron Microscopy.

Results: We obtained 450 Calcein+ and CD41+ platelets per megakaryocyte which is the highest value through the microfluidic system at 600 µl/min (shear stress between 0.042 Pa and 97 Pa). These platelets have a well-defined cell membrane and organelles including alpha- and dense granules, and open canalicular systems.

Conclusion(s): We propose the use of microfluidic systems, particularly those that mimic pulmonary microvasculature, as a viable method for platelet biogenesis. The quantity and quality of the generated platelets demonstrate that they would be viable candidates for in vitro platelet biogenesis platforms and compare favourably with other systems previously reported in the literature.

Learning Objectives:

Upon completion, participant will be able to apply the methods for platelet generation by using in vitro systems.
Upon completion, participant will be able to observe the different outcomes between two distinct channel designs.
Upon completion, participant will be able to read the experimental results.