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Mouse and human HSPC immobilization in liquid culture by CD43 or CD44-antibody coating

Dirk Loeffler, Weijia Wang, Alois Hopf, Oliver Hilsenbeck, Paul E. Bourgine, Fabian Rudolf, Ivan Martin and Timm Schroeder

Key points

  • Anti-CD43 and -CD44 antibody coating immobilizes live mouse and human hematopoietic stem and progenitor cells.

  • This enables 2D colony formation, medium exchange without cell identification loss, and increased throughput of time-lapse imaging.

Abstract

Keeping track of individual cell identifications is imperative to the study of dynamic single cell behavior over time. Highly motile hematopoietic stem and progenitor cells (HSPCs) migrate quickly and do not adhere, and thus must be imaged very frequently to keep cell identifications. Even worse, they are also flushed away during medium exchange. To overcome these limitations, we tested antibody coating for reducing HSPC motility in vitro. Anti-CD43 and -CD44 antibody coating reduced cell motility of mouse and human HSPCs in a concentration dependent manner. This enables 2D colony formation without cell mixing in liquid cultures, massively increases time-lapse imaging throughput, and maintains cell positions also during media exchange. Anti-CD43, but not -CD44 coating reduces mouse HSPC proliferation with increasing concentrations. No relevant effects on cell survival or myeloid and megakaryocyte differentiation of hematopoietic stem cells and multipotent progenitors 1-5 (MPPs) were detected. Human umbilical cord hematopoietic CD34+ cell survival, proliferation and differentiation was not affected by either coating. This approach both, massively simplifies and accelerates continuous analysis of suspension cells, and enables the study of their behavior in dynamic rather than static culture conditions over time.

  • Submitted July 31, 2017.
  • Accepted January 13, 2018.