Details
| Original language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Pages | 71-84 |
| Number of pages | 14 |
| ISBN (electronic) | 978-1-4939-9574-5 |
| Publication status | Published - 2019 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 2017 |
| ISSN (Print) | 1064-3745 |
| ISSN (electronic) | 1940-6029 |
Abstract
Cell adhesion is implicated in many physiological settings such as the retention of hematopoietic stem cells (HSCs) in their bone marrow niches or their migration into the bloodstream. During HSC mobilization these adhesion sites are cleaved and have to be newly formed during HSC homing and engraftment. To determine the adhesive properties of HSCs on different extracellular matrix (ECM) molecules, we present a microfluidic shear force assay, where a laminar flow is used to detach a semi-adherent cell population, the HSC model cell line KG-1a, from an ECM protein-coated substrate. This technique combines the high throughput of population-based assays with the ability to observe cell detachment in real time. Additionally, it is suitable for weakly adherent cells, as the setup allows cell incubation on various substrates and application of shear stress ranging several orders of magnitude in one setup without additional washing or transfer steps. As a measure for the adhesion strength of the studied cell population on the substrate, the critical shear force τ50 is determined which is required to remove 50% of the initially adherent cell fraction.
Keywords
- Cell adhesion, Leukemic cells, Microfluidic assay, Protein coating, Shear force
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
Cite this
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Methods in Molecular Biology. 2019. p. 71-84 (Methods in Molecular Biology; Vol. 2017).
Research output: Chapter in book/report/conference proceeding › Contribution to book/anthology › Research › peer review
}
TY - CHAP
T1 - Microfluidic Shear Force Assay to Determine Cell Adhesion Forces
AU - Hümmer, Julia
AU - Koc, Julian
AU - Rosenhahn, Axel
AU - Lee-Thedieck, Cornelia
PY - 2019
Y1 - 2019
N2 - Cell adhesion is implicated in many physiological settings such as the retention of hematopoietic stem cells (HSCs) in their bone marrow niches or their migration into the bloodstream. During HSC mobilization these adhesion sites are cleaved and have to be newly formed during HSC homing and engraftment. To determine the adhesive properties of HSCs on different extracellular matrix (ECM) molecules, we present a microfluidic shear force assay, where a laminar flow is used to detach a semi-adherent cell population, the HSC model cell line KG-1a, from an ECM protein-coated substrate. This technique combines the high throughput of population-based assays with the ability to observe cell detachment in real time. Additionally, it is suitable for weakly adherent cells, as the setup allows cell incubation on various substrates and application of shear stress ranging several orders of magnitude in one setup without additional washing or transfer steps. As a measure for the adhesion strength of the studied cell population on the substrate, the critical shear force τ50 is determined which is required to remove 50% of the initially adherent cell fraction.
AB - Cell adhesion is implicated in many physiological settings such as the retention of hematopoietic stem cells (HSCs) in their bone marrow niches or their migration into the bloodstream. During HSC mobilization these adhesion sites are cleaved and have to be newly formed during HSC homing and engraftment. To determine the adhesive properties of HSCs on different extracellular matrix (ECM) molecules, we present a microfluidic shear force assay, where a laminar flow is used to detach a semi-adherent cell population, the HSC model cell line KG-1a, from an ECM protein-coated substrate. This technique combines the high throughput of population-based assays with the ability to observe cell detachment in real time. Additionally, it is suitable for weakly adherent cells, as the setup allows cell incubation on various substrates and application of shear stress ranging several orders of magnitude in one setup without additional washing or transfer steps. As a measure for the adhesion strength of the studied cell population on the substrate, the critical shear force τ50 is determined which is required to remove 50% of the initially adherent cell fraction.
KW - Cell adhesion
KW - Leukemic cells
KW - Microfluidic assay
KW - Protein coating
KW - Shear force
UR - http://www.scopus.com/inward/record.url?scp=85067522846&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9574-5_6
DO - 10.1007/978-1-4939-9574-5_6
M3 - Contribution to book/anthology
C2 - 31197769
AN - SCOPUS:85067522846
SN - 978-1-4939-9573-8
T3 - Methods in Molecular Biology
SP - 71
EP - 84
BT - Methods in Molecular Biology
ER -