Effect of Different Trypsin Concentrations on SMCC-7721 Cell Adhesion

Jin, Yan; Tian, Liguo; Xie, Chenchen; Liu, Yan; Lu, Zhengcheng; Wang, Zuobin

doi:10.18178/ijpmbs.10.3.103-107

Cited by 2 publications

(1 citation statement)

References 3 publications

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“…Trypsin is a serine protease that is near-universally used to detach cells from culture substrates during routine cell culture and passaging. Depending on the concentration of trypsin used [40], trypsin can detach cells on the timescale of minutes to tens of minutes. We thus propose and demonstrate the simultaneous employment of shear flow and trypsin treatment to assess the cell adhesion strength of a strongly adherent cell type and elucidate the type of data that can be obtained from such an assay.…”

Section: Introductionmentioning

confidence: 99%

Integrating Shear Flow and Trypsin Treatment to Assess Cell Adhesion Strength

Patel,

Bhavanam,

Keenan

et al. 2023

Preprint

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Cell adhesion is of fundamental importance in cell and tissue organization, and for designing cell-laden constructs for tissue engineering. Prior methods to assess cell adhesion strength for strongly adherent cells using hydrodynamic shear flow either involved the use of specialized flow devices to generate high shear stress or used simpler implementations like larger height parallel plate chambers that enable multi-hour cell culture but generate low shear stress and are hence more applicable for weakly adherent cells. Here, we propose a shear flow assay for adhesion strength assessment of strongly adherent cells that employs off-the-shelf parallel plate chambers for shear flow as well as simultaneous trypsin treatment to tune down the adhesion strength of cells. We implement the assay with a strongly adherent cell type and show that shear stress in the 0.07 to 7 Pa range is sufficient to dislodge the cells with simultaneous trypsin treatment. Imaging of cells over a square centimeter area allows cell morphological analysis of hundreds of cells. We show that the cell area of cells that are dislodged, on average, does not monotonically increase with shear stress at the higher end of shear stresses used and suggest that this can be explained by the likely higher resistance of high circularity cells to trypsin digestion. The adhesion strength assay proposed can be easily adapted by labs to assess the adhesion strength of both weakly and strongly adherent cell types and has the potential to be adapted for substrate stiffness-dependent adhesion strength assessment in mechanobiology studies.

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Section: Introductionmentioning

confidence: 99%

Integrating Shear Flow and Trypsin Treatment to Assess Cell Adhesion Strength

Patel,

Bhavanam,

Keenan

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Integrating shear flow and trypsin treatment to assess cell adhesion strength

Patel,

Bhavanam,

Keenan

et al. 2023

Biointerphases

View full text Add to dashboard Cite

Cell adhesion is of fundamental importance in cell and tissue organization and for designing cell-laden constructs for tissue engineering. Prior methods to assess cell adhesion strength for strongly adherent cells using hydrodynamic shear flow either involved the use of specialized flow devices to generate high shear stress or used simpler implementations like larger height parallel plate chambers that enable multihour cell culture but generate low wall shear stress and are, hence, more applicable for weakly adherent cells. Here, we propose a shear flow assay for adhesion strength assessment of strongly adherent cells that employs off-the-shelf parallel plate chambers for shear flow as well as simultaneous trypsin treatment to tune down the adhesion strength of cells. We implement the assay with a strongly adherent cell type and show that wall shear stress in the 0.07–7 Pa range is sufficient to dislodge the cells with simultaneous trypsin treatment. Imaging of cells over a square centimeter area allows cell morphological analysis of hundreds of cells. We show that the cell area of cells that are dislodged, on average, does not monotonically increase with wall shear stress at the higher end of wall shear stresses used and suggest that this can be explained by the likely higher resistance of high circularity cells to trypsin digestion. The adhesion strength assay proposed can be used to assess the adhesion strength of both weakly and strongly adherent cell types and has the potential to be adapted for substrate stiffness-dependent adhesion strength assessment in mechanobiology studies.

show abstract

Effect of Different Trypsin Concentrations on SMCC-7721 Cell Adhesion

Cited by 2 publications

References 3 publications

Integrating Shear Flow and Trypsin Treatment to Assess Cell Adhesion Strength

Integrating Shear Flow and Trypsin Treatment to Assess Cell Adhesion Strength

Integrating shear flow and trypsin treatment to assess cell adhesion strength

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