Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture

Kerwin Kwek Zeming, Yuko Sato, Lu Yin, Nai Jia Huang, Lan Hiong Wong, Hooi Linn Loo, Ying Bena Lim, Chwee Teck Lim, Jianzhu Chen, Peter R. Preiser, Jongyoon Han

研究成果: Article査読

7 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)3445-3460
ページ数16
ジャーナルLab on a Chip
20
18
DOI
出版ステータスPublished - 2020 9月 21

ASJC Scopus subject areas

  • バイオエンジニアリング
  • 生化学
  • 化学 (全般)
  • 生体医工学

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