High-throughput assessment of mechanical properties of stem cell derived red blood cells, toward cellular downstream processing

Scientific Reports
Ewa GuzniczakHelen Bridle

Abstract

Stem cell products, including manufactured red blood cells, require efficient sorting and purification methods to remove components potentially harmful for clinical application. However, standard approaches for cellular downstream processing rely on the use of specific and expensive labels (e.g. FACS or MACS). Techniques relying on inherent mechanical and physical properties of cells offer high-throughput scalable alternatives but knowledge of the mechanical phenotype is required. Here, we characterized for the first time deformability and size changes in CD34+ cells, and expelled nuclei, during their differentiation process into red blood cells at days 11, 14, 18 and 21, using Real-Time Deformability Cytometry (RT-DC) and Atomic Force Microscopy (AFM). We found significant differences (p < 0.0001; standardised mixed model) between the deformability of nucleated and enucleated cells, while they remain within the same size range. Expelled nuclei are smaller thus could be removed by size-based separation. An average Young's elastic modulus was measured for nucleated cells, enucleated cells and nuclei (day 14) of 1.04 ± 0.47 kPa, 0.53 ± 0.12 kPa and 7.06 ± 4.07 kPa respectively. Our identification and quantification of significant...Continue Reading

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Citations

Jan 10, 2020·Lab on a Chip·Ewa GuzniczakHelen Bridle
Oct 8, 2019·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Ricardo H PiresOliver Otto
Mar 1, 2020·Cells·Ruixue ZhuIgor Meglinski
Aug 15, 2020·Lab on a Chip·Kerwin Kwek ZemingJongyoon Han

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Methods Mentioned

BETA
Magnetic Activated Cell Sorting
FACS
atomic force microscopy
AFM
MDA
Knockout
flow cytometry
chip

Software Mentioned

ShapeOut
MatLab
Huygens Professional
GraphPad Prism
spm
FlowJo
JPK

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