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A Microcavity Array-Based 3D Model System of the Hematopoietic Stem Cell Niche

  • Eric GottwaldEmail author
  • Cordula Nies
  • Patrick Wuchter
  • Rainer Saffrich
  • Roman Truckenmüller
  • Stefan Giselbrecht
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2017)

Abstract

Despite huge advances in recent years, the interaction between hematopoietic stem and progenitor cells (HSPCs) and their niches in the bone marrow is still far from being fully understood. One reason is that hematopoiesis is a multi-step maturation process leading to HSPC heterogeneity. Subpopulations of HSPCs can be identified by clonogenic assays or in serial transplantation experiments in mice following sublethal irradiation, but it is very complex to reproduce or even maintain stem cell plasticity in vitro. Advanced model systems have been developed that allow to precisely control and analyze key components of the physiologic microenvironment for not only fundamental research purposes but, as a long-term goal, also for clinical applications. In this chapter, we describe our approach of building an artificial hematopoietic stem cell niche in the form of polymer film-based microcavities with a diameter of 300 μm and a depth of up to 300 μm and arranged in a 634-cavity array. The polymer films are provided with 3 μm pores and thus allow perfusion of the culture medium. The microcavity arrays can be inserted into a microbioreactor where a closed circulation loop can be tightly controlled with regard to medium flow and gas supply. The microcavity arrays were used for a three-dimensional (3D) co-culture of MSCs and HSPCs in a defined ratio over a time period of up to 21 days. With this setup, it could be demonstrated that the HSPCs maintained their stem cell characteristics more efficiently as compared to conventional monolayer co-culture controls.

Key words

Microthermoforming Microcavity array MSC-HSPC co-culture Perfusion Microbioreactor 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Eric Gottwald
    • 1
    Email author
  • Cordula Nies
    • 1
  • Patrick Wuchter
    • 2
    • 3
  • Rainer Saffrich
    • 4
    • 3
    • 5
  • Roman Truckenmüller
    • 6
  • Stefan Giselbrecht
    • 6
  1. 1.Institute of Functional InterfacesKarlsruhe Institute of TechnologyEggenstein-LeopoldshafenGermany
  2. 2.Institute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, Heidelberg UniversityMannheimGermany
  3. 3.German Red Cross Blood Service Baden-Württemberg — HessenMaastrichtThe Netherlands
  4. 4.Institute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, Heidelberg UniversityMannheimGermany
  5. 5.Department of Medicine V (Hematology, Oncology, Rheumatology)Heidelberg UniversityHeidelbergGermany
  6. 6.Department of Complex Tissue Regeneration, MERLN Institute for Technology-Inspired Regenerative MedicineMaastricht UniversityMaastrichtThe Netherlands

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