细胞功能测定

In this chapter, we describe the derivation and characterization of nine hIn this chapter, we describe the derivation and characterization of nine human embryonic stem cells (hESC) (VAL-3 to -11B) from different developmental embryo stages (inner cell mass from a blastocyst, morula, and b ...

The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quant ...

Although since 1998 more than 1,200 different hESC lines have been established worldwide, there is still a recognized interest in the establishment of new lines of hESC, particularly from HLA types and ethnic groups underrepresented among the currently available lines. The methodology ...

The analysis of DNA-methylation on a genome-wide scale by next-generation sequencing techniques is an invaluable tool towards the understanding of the epigenetic basis of cellular differentiation. Methylated DNA immunoprecipitation (MeDIP) is an immunocapturing method u ...

Chromatin immunoprecipitation (ChIP) followed by microarray-based (ChIP-Chip) or next-generation sequencing-based (ChIP-Seq) analysis has been established as a powerful and widely used method to investigate DNA–protein interactions relative to a genomic location in vi ...

During culture adaptation, human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) tend to acquire chromosomal aberrations. Generally, stem cell lines are screened for large-scale chromosomal changes using low resolution karyotype analysis. Recent stud ...

Neural differentiation of human embryonic (ES) and induced pluripotent (iPS) stem cell lines has been used for research in early human development, drug discovery, and cell replacement therapies. It is critical to establish generic differentiation protocols to compare the neural sp ...

The differentiation of human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs) towards functional neurons particularly hold great potential for the cell-based replacement therapy in neurodegenerative diseases. Here, we describe a stepwise dif ...

Embryonic stem cells (ESCs) are unique cells, which have the ability to differentiate into all cell types that comprise the adult organism. Furthermore, ESCs can infinitely self-renew under optimized conditions. These features place human ESCs (hESCs) in a position where these cells can be ...

Human embryonic and induced pluripotent stem cells (hESCs and hiPSCs) offer a platform technology with the potential for developmental biology and cell-based therapy. Therefore, robust and cost-effective ways for mass production of them is necessary. Here, we have presented a protoc ...

Experimental manipulation of hESCs has been hampered by their fragility and susceptibility to apoptosis when dissociated into single cells. The OxF lines are particularly robust and may be successfully passaged as single cells, with the inclusion of ROCK inhibitor in the medium. The pro ...

Human embryonic stem cells (HESCs) are of great interest in biology and medicine due to their ability to grow indefinitely in culture while maintaining their ability to differentiate into all different cell types in the human body. In addition, HESCs can be used for better understanding the key d ...

Human embryonic stem cells (hESCs) are an invaluable cell source to study human embryogenesis and development and for exploring the nature of human diseases. Moreover, hESCs can serve as an unlimited source of cells for cell therapy. The first hESC lines were derived from frozen blastocyst-s ...

Gene silencing approaches afford investigators the ability to gain important insight into the normal functional requirements of specific epidermal proteins and promise to yield a powerful therapeutic means to dampen the level of proteins that are mutated or frequently overexpre ...

Detection of in vitro protein–DNA interaction is one of many investigational analyses for transcription factor regulation of gene promoters. The electrophoretic mobility shift assay (EMSA) has proven widely popular in this respect by integrating individual techniques (prot ...

Protein–DNA interactions, such as those that are necessary for transcription, are critical in regulating cellular function and behavior. The identification of DNA sequences that interact with transcriptional regulatory proteins is an important step necessary to better under ...

DNA microarray technology is a powerful tool for genome-wide gene expression analysis of biological samples including skin and epidermal keratinocytes. Here we review the methodology for expression profiling analysis of skin tissue or purified keratinocytes from mice. We expla ...

Epithelial tissues exhibit optimal conditions for studying cellular differentiation since the differentiation status of a single cell can be determined by its distance to the basal membrane. For that reason Laser Capture Microdissection (LCM) may serve as a perfect tool to compare the c ...

Because of its accessibility, skin has been among the first organs analyzed using DNA microarrays; psoriasis, melanomas, carcinomas, chronic wound biopsies, and epidermal keratinocytes in culture have been intensely investigated. Skin has everything: stem cells, differentia ...

The mouse mammary gland is a unique organ since although the mammary gland primordium forms during embryogenesis, the majority of development occurs postnatally upon hormonal stimulation at puberty and full functionality (i.e. lactation) is not achieved until after parturition. S ...