Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture

In vitro erythroid cultures from human hematopoietic stem cells produce immature red blood cells (RBCs) called reticulocytes, which are important for RBCs production, and are widely used in scientific studies of malaria pathology, hematological diseases and protein translation. However, in vitro ret...

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Bibliographic Details
Published in:Lab on a chip 2020-09, Vol.2 (18), p.3445-346
Main Authors: Zeming, Kerwin Kwek, Sato, Yuko, Yin, Lu, Huang, Nai-Jia, Wong, Lan Hiong, Loo, Hooi Linn, Lim, Ying Bena, Lim, Chwee Teck, Chen, Jianzhu, Preiser, Peter R, Han, Jongyoon
Format: Article
Language:English
Subjects:
Bioprocessing
Culture
Deformation
Erythrocytes
Fluorescence
Formability
Fractionation
Lateral displacement
Malaria
Microfluidics
Nuclei (cytology)
Production costs
Purity
Recovery
Reticulocytes
Stem cells
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Summary:In vitro erythroid cultures from human hematopoietic stem cells produce immature red blood cells (RBCs) called reticulocytes, which are important for RBCs production, and are widely used in scientific studies of malaria pathology, hematological diseases and protein translation. However, in vitro reticulocyte cultures contain expelled cell nuclei and erythroblasts as undesirable by-products and current purification methods such as density gradient centrifugation and fluorescence-activated cell sorting (FACS) are not optimal for integrated bioprocessing and downstream therapeutic applications. Developments in Dean flow fractionation (DFF) and deterministic lateral displacement (DLD) microfluidic sorting methods are ideal alternatives due to label-free size sorting, throughput scalability and low manufacturing cost. DFF sorting of reticulocytes from whole erythroid culture showed a 2.4-fold increase in cell recovery compared to FACS albeit with a lower purity; DLD sorting showed comparable cell recovery and purity with FACS using an inverse-L pillar structure to emphasize size and deformability sorting of reticulocytes. The viability and functional assurance of purified reticulocytes showed conserved cell deformability and supported the propagation of malaria parasites. Collectively, our study on label-free RBCs isolation represents a significant technical advancement towards developing in vitro generated viable human RBCs, opening opportunities for close-loop cell manufacturing, downstream therapeutic and research purposes. Developments in Dean flow fractionation (DFF) and deterministic lateral displacement (DLD) for label-free purification of cultured RBCs from human hematopoietic stem cells. An advancement in sorting and closed-loop manufacturing of viable human RBCs.
ISSN:1473-0197
1473-0189
DOI:10.1039/c9lc01128e