The assessment of viable osteocytes within bone tissue is of crucial importance. Osteocytes are the most abundant cells in bone. Due to their interconnectivity in the bone matrix they are hypothesised to play an important role in the maintenance of the extracellular matrix of bone. The death of o ...
In tissue engineering, a variety of methods are commonly used to evaluate survival of cells inside tissues or three-dimensional (3D) carriers. Among these methods confocal laser scanning microscopy opened accessibility of 3D tissue using live cell imaging into the tissue or 3D scaffold ...
Quantification of cell viability and distribution within engineered tissues currently relies on representative histology, phenotypic assays, and destructive assays of viability. To evaluate uniformity of cell density throughout 3D collagen scaffolds prior to in vivo use, a no ...
Testing the effects of compounds on the viability of cells grown in culture is widely used as a predictor of potential toxic effects in whole animals. Among the several alternative assays available, measuring the levels of ATP is the most sensitive, reliable, and convenient method for monitori ...
Today, obtaining mechanistic insights into biological, toxicological, and pathological processes is of upmost importance. Researchers aim to obtain as many as possible data from one cell sample to understand the biological processes under study. Multiplexing, which is the ability ...
The gross majority of classical apoptotic hallmarks can be rapidly examined by multiparameter flow cytometry. As a result, cytometry became a technology of choice in diverse studies of cellular demise. In this context, a novel class of substituted unsymmetrical cyanine SYTO probes has re ...
Caspases are critical regulators of the apoptotic program, responsible for the harmonic dismantling of the cell. Cell death can occur by way of different options (necroptosis, necrosis, extreme autophagy) but once caspases are fully engaged it will take the apoptotic route. Hence, in gene ...
Annexin V/7-amino-actinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Early apoptotic cells express phosphatidylserines (PS) on the outer leaflet of the plasma membrane. PS can be stained by labeled annexin V. Late apoptotic cel ...
A variety of assays, and rationales for their use, exist to monitor viability and/or survival following cellular exposure to insult. Two commonly used in vitro assays are the sulforhodamine B assay and the clonogenic survival assay which can be used to monitor the efficacy of anticancer agents, ...
We describe here the use of the xCELLigence system for label-free and real-time monitoring of cell �viability. The xCELLigence system uses specially designed microtiter plates containing interdigitated gold microelectrodes to noninvasively monitor the viability of cultur ...
The Alamar Blue assay is based on enzymatic reduction of indicator dye by viable cells and serves as an effective tool for assessing cell proliferation and as a screening technique. It can be applied in studies concentrating on animal, plant, yeast, and bacteria cells. Among the various methods for ...
WST-8 is one of the newer generation formazan-based dyes, which release the converted product into the medium in a soluble form. This allows for a non-destructive determination of viability enabling the cells to be subject to further investigations. This is a major advantage in cases where cell p ...
The MTT reduction assay is used to determine the level of metabolic activity in eukaryotic cells, �including animal, plant, and fungal cells. If the metabolic rate is constant, the technique can be employed to count living cells in a sample. Once it is set up, the method is very robust, and can be automatized to ...
One of the traditional methods of cell viability analysis is the use of trypan blue dye exclusion staining. This technique has been the standard methodology used in academic research laboratories and industrial biotechnology plants. Cells were routinely counted manually with a hemoc ...
The quantification of live and dead cells in a substrate is often an essential step in cell biology research. A staining protocol that acts differently on live and on dead cells is applied and the number of cells visible is counted using a microscope. Often this counting is done manually or only evaluated ...
Many existing protocols for neuronal differentiation of human pluripotent cells result in heterogeneous cell populations and unsynchronized differentiation, necessitating the development of methods for labeling specific cell populations. Here we describe how microR ...
Heterogeneity of stem cell populations is a well-known but poorly characterized phenomenon. Here, we demonstrate the qualitative and quantitative power of single-cell transcript analysis to characterize transcriptome dynamics in embryonic stem cells (ESC). In this chapter, we ...
In this chapter, we describe an effective and reproducible protocol for neural differentiation of human pluripotent stem cells in three dimensional (3D) collagen and MartigelTM gels. We have used this protocol to generate embryoid bodies (EBs) from dissociated suspension cultures of ...
Human embryonic stem cells (hESCs) are pluripotent cells that have the potential to be used for tissue engineering and regenerative medicine. Biochemical and biological agents are widely used to induce hESC differentiation. However, it would be better if we could induce the differentia ...
Human embryonic stem cells (hESCs) have the capacity to differentiate in vivo and in vitro into cells from all three germ lineages. The in vitro generation of lung cells and tissues from hESCs creates opportunities for fundamental research, drug development or cell-replacement therapy. In ...