ASTM F3088-22

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Designation: F3088 − 14 F3088 − 22
Standard Test Method Practice for
Use of a Centrifugation Method to Quantify/Study Cell-
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Material Adhesive Interactions
This standard is issued under the fixed designation F3088; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This test method covers a centrifugation cell adhesion assay that can be used to detect changes in adhesive characteristics of
cells with passage or treatments. This approach measures the force required to detach cells from a substrate. Adhesion, among
many variables, may vary due to changes in the phenotype of the cells.
1.2 This test method does not cover methods to verify the uniformity of coating of surfaces, nor does it cover methods for
characterizing surfaces.
1.3 The cells may include adult, progenitor or stem cells from any species. The types of cells may include chondrocytes,
fibroblasts, osteoblast, islet cells, or other relevant adherent cell types.
1.4 This test method does not cover methods for isolating or harvesting of cells. This test method does not cover test methods to
quantitate changes in gene expression, or changes in biomarker type or concentration, as identified by immunostaining. Nor does
this test method cover quantitative image analysis techniques.
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility
of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory
limitations prior to use.
2. Referenced Documents
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2.1 ASTM Standards:
F2603 Guide for Interpreting Images of Polymeric Tissue Scaffolds
F2664 Guide for Assessing the Attachment of Cells to Biomaterial Surfaces by Physical Methods
F2739 Guide for Quantifying Cell Viability and Related Attributes within Biomaterial Scaffolds
F2944 Practice for Automated Colony Forming Unit (CFU) Assays—Image Acquisition and Analysis Method for Enumerating
and Characterizing Cells and Colonies in Culture
3. Summary of Test Method
3.1 Centrifugation Cell Assay on Cell Populations—A conventional centrifuge can be used to apply a normal or shear force to cells
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This test method practice is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devices and is the direct responsibility of
Subcommittee F04.43 on Cells and Tissue Engineered Constructs for TEMPs.
Current edition approved May 1, 2014April 1, 2022. Published June 2014May 2022. Originally approved in 2014. Last previous edition approved in 2014 as F3088 – 14.
DOI: 10.1520/F3088-14.10.1520/F3088-22.
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For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
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F3088 − 22
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depending on the orientation of the cells with respect to the centrifugal force (1-3). The centrifugal force that the cells are subject
to can be calculated according to the following formula assuming that the plates are 90° (normal) to the spindle of the centrifuge.
F 5 p 2 p V RCF (1)
~ !
D cell medium cell
where:
F = detachment force applied per cell,
D
V = cell volume,
cell
p = density of the cell,
cell
p = the density of the surrounding medium, and
medium
2
RCF = Relative Centrifugal Forces = rῳ , where r = centrifugation radius and ῳ = centrifugation speed.
See X1.6 for example of a calculation of “detachment force per cell.”
3.1.1 Such tests are easy to conduct and the results represent a population average. The method can be performed in a moderately
outfitted cell biology lab with an inverted microscope or a fluorescence microplate reader and a centrifuge with a microtiter plate
holder. Three factors need to be considered when using this methodology: the potential influences of forces applied during the
period of spin up time, the maximum spin speed, and the test duration. The method is only able to correlate with cell detachment
with the maximum force applied after the centrifuge has reached its set spin speed. It should also be consi
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