细胞功能测定

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 ...

Hepatocytes derived from human embryonic stem (hES) cells are a potential cell source for regenerative medicine. However, the successful differentiation of hES cells into mature hepatocytes has been difficult to achieve because the definitive mechanisms governing hepatocyte ...

Efficient hepatic differentiation technologies designed for patient specific induced pluripotent stem (iPS) cells may provide an unlimited hepatocyte source which can be utilized in drug screening, disease modeling, and cell therapy. This chapter describes the methods we use to d ...

The human liver is a vital organ within the body and plays a major role in normal homeostasis. The “work horse” of the liver, termed the “hepatocyte,” is estimated to make up approximately 70–80% of the liver’s mass. Therefore, the study of hepatocyte biology has an important role to play in medicine and the drug ...

Human adult cartilage has limited capacity for self-renewal. Accordingly, repair of cartilage tissue damaged as a result of acute traumatic injury or via chronic wear or degenerative disease, such as arthritis, is a major clinical problem. Human embryonic stem cells (hESCs) could provide ...

Human induced pluripotent stem (iPS) cells are promising sources of disease modeling and regenerative medicine. However, differentiation properties of human iPS cells to specific lineages still remain to be fully clarified. Here, we describe a protocol for differentiating human i ...

Human embryonic stem cells (hESCs) and patient-specific human induced pluripotent stem cells (hiPSCs) are valuable reagents for studying the earliest stages of hematopoietic genesis and for modeling the developmental basis of hematologic disorders, and they may also have the pote ...

The establishment of human embryonic stem cell (hESC) lines, as well as the recent induced pluripotent stem cells (hiPSC), has greatly expanded our knowledge about the early development in human ontogeny. In the past decade, hESCs and hiPSCs have been proven excellent tools in characterizat ...