A novel time resolved fluorometric assay of anoikis using Europium-labelled Annexin V in cultured adherent cells

Engbers-Buijtenhuijs, P.; Kamphuis, Marloes M. J.; Veer, G. van der Sluijs; Haanen, C.; Poot, Andreas A.; Feijén, Jan; Vermes, I.

doi:10.1007/s10495-005-0816-4

Cited by 14 publications

(9 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Harumi Miyagi et al ( 31 ) observed donor-to-donor variation of the expression of extracellular matrix proteins with different human dental pulp stem cells from deciduous teeth, which could justify the different adhesion behavior between the cultures presented in Figure 6 . This is in agreement with the fact that mesenchymal stem cells, as anchorage-dependent cells, can undergo cell death by the lack of appropriated attachment to a substrate ( 32 ).…”

Section: Resultssupporting

confidence: 87%

Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion

Paim

Braghirolli

Cardozo

et al. 2018

Braz J Med Biol Res

View full text Add to dashboard Cite

Cell adhesion in three-dimensional scaffolds plays a key role in tissue development. However, stem cell behavior in electrospun scaffolds under perfusion is not fully understood. Thus, an investigation was made on the effect of flow rate and shear stress, adhesion time, and seeding density under direct perfusion in polycaprolactone electrospun scaffolds on human dental pulp stem cell detachment. Polycaprolactone scaffolds were electrospun using a solvent mixture of chloroform and methanol. The viable cell number was determined at each tested condition. Cell morphology was analyzed by confocal microscopy after various incubation times for static cell adhesion with a high seeding density. Scanning electron microscopy images were obtained before and after perfusion for the highest flow rate tested. The wall pore shear stress was calculated for all tested flow rates (0.005–3 mL/min). An inversely proportional relationship between adhesion time with cell detachment under perfusion was observed. Lower flow rates and lower seeding densities reduced the drag of cells by shear stress. However, there was an operational limit for the lowest flow rate that can be used without compromising cell viability, indicating that a flow rate of 0.05 mL/min might be more suitable for the tested cell culture in electrospun scaffolds under direct perfusion.

show abstract

Section: Resultssupporting

confidence: 87%

Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion

Paim

Braghirolli

Cardozo

et al. 2018

Braz J Med Biol Res

View full text Add to dashboard Cite

show abstract

“…This protocol is optimized for cells in suspension and not for adherent cells. If adherent cells must be used, trypsinization should be avoided or at least minimized if possible to prevent anoikis-induced cell death [28, 29]. Although trypsinization usually does not affect traditional cell death assays (e.g., MTT, XTT assays), it can affect proteins bound to the cellular membrane and change the permeability profile of cells.…”

Section: Methodsmentioning

confidence: 99%

Flow Cytometry-Based Detection and Analysis of BCL-2 Family Proteins and Mitochondrial Outer Membrane Permeabilization (MOMP)

Ludwig

Maxcy

LaBelle

2018

Methods in Molecular Biology

View full text Add to dashboard Cite

The BCL-2 family of proteins orchestrates a complex signaling network that governs the balance between cellular survival and death. A comprehensive understanding of the mechanistic interactions between these proteins continues to evolve in normal and malignant cells. The functional variation by individual BCL-2 proteins in different cell types has driven clinical therapeutic development in targeting individual BCL-2 members with the goal of fine-tuning cell death in diseased cells. Given the importance of understanding and validating the effect of activating or inhibiting BCL-2 protein interactions in individual cells, the methods used to measure apoptotic cell death have undergone increased scrutiny. Here, we describe two in vitro flow cytometry-based methods that are useful in measuring BCL-2 proteins and mitochondrial-based cell death in complex cell populations.

show abstract

“…15,16 In both cases, the recognition agent is coupled to a second part which can be detected (optical detection, radio labeling, magnetic particles for cell sorting, immunodetection and etc.). 9,10,17,18 Large numbers of the annexin-superfamily proteins are reported to be expressed in eukaryotes. 19 Annexins are widespread, greatly conserved and mostly intracellular proteins that generally are distributed in tissues.…”

Section: Molecular Structure Of Annexin and Detection Of Apoptosismentioning

confidence: 99%

Comparison of different probes based on labeled annexin V for detection of apoptosis

2014

View full text Add to dashboard Cite

show abstract

A novel time resolved fluorometric assay of anoikis using Europium-labelled Annexin V in cultured adherent cells

Abstract: The novel time resolved assay provides the ability to analyse the cell death cascade in adherent cell cultures of the same sample at the same time in a sensitive and reproducible way.

Cited by 14 publications

References 27 publications

Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion

Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion

Flow Cytometry-Based Detection and Analysis of BCL-2 Family Proteins and Mitochondrial Outer Membrane Permeabilization (MOMP)

Comparison of different probes based on labeled annexin V for detection of apoptosis

Contact Info

Product

Resources

About