细胞功能测定

Thin-layer chromatography (TLC) is a technique that has been routinely used for the separation and identification of lipids. Here we describe an optimized protocol for the steady state labeling, separation, and quantification of yeast phospholipids using 1-D TLC.

Lipid analysis performed by nano-electrospray ionization mass spectrometry is a highly sensitive method for quantification of lipids including all lipid species of a given lipid class. Various instrumental setups are used for quantitative lipid analysis, including different ...

Isolation and culture of primary embryonic stem (ES) cell colonies are the first critical step towards establishment of stable ES cell lines. Here we introduce a novel method designated as “Separate and Seed” that contributes remarkably to efficient derivation of bovine primary ES cell co ...

Regulation of early development can be directly interrogated in the embryo or can be studied in cultured cells isolated in the laboratory. New understanding of the developmental stages and the signalling requirements of the pig embryo have enabled the development of improved protocols ...

Despite their agricultural and biomedical importance, embryonic stem cells (ESCs) are yet to be isolated for the pig or the domestic ungulates in general. This suggests that methods which have been used successfully in mice may not be applicable to these. In this chapter we describe a new method for t ...

The derivation of canine embryonic stem cells (cESCs) represents a significant achievement and opens the door to further stem cell research and therapies in the dog. Canines share a common environment with humans and exhibit a host of genetic diseases, many of which have human parallels. Thus, t ...

Embryonic stem (ES) cells are derived from the inner cell masses of preimplantation embryos. ES cells are pluripotent cells with the capacity for long-term propagation and broad differentiation plasticity. These cells have an exceptional functional feature in that they can differen ...

Mammalian stem cells are invaluable research resources for the study of cell and embryonic development as well as practical tools for use in the production of genetically engineered animals and further therapeutics. It is important that we further our knowledge and understanding of a var ...

The derivation of mouse embryonic stem cells (mESCs) from the inner cell mass (ICM) of mouse preimplantation blastocysts has provided tremendous opportunities in exploring mammalian development and pluripotent stem cell biology (Martin, Proc Natl Acad Sci U S A 78(12):7634–7638, 1981; ...

Mouse postimplantation epiblast cultured in activin and basic fibroblast growth factor gives rise to continuously growing epiblast stem cells (EpiSCs) that share key properties with postimplantation epiblast, such as DNA methylation and an inactive X-chromosome. EpiSCs also s ...

Mouse epiblast stem cells (EpiSCs) are pluripotent embryonic cells that can be used to interrogate developmental transitions that occur during gastrulation. EpiSCs can also be robustly differentiated into functional somatic and germ cell derivatives making them a useful tool for s ...

Induction of neural differentiation from embryonic pluripotent stem cells (ES/EpiSc), both of mouse and primates, has been extensively published by several research teams. However, direct derivation of organized neuroectoderm in vitro from blastocyst stage embryos of rodents or ...

The isolation and culture of neural progenitor cells (NPCs) from pluripotent stem cells has facilitated in vitro mechanistic studies of diseases related to the nervous system, as well as discovery of new medicine. In addition, NPCs are envisioned to play a crucial role in future cell replaceme ...

A group of pluripotent cells appearing during mammalian embryogenesis is the source for all the cell lineages that compose the embryo proper. In mice, pluripotent cells are first established in the inner cell mass (ICM) of the preimplantation blastocyst. After implantation, the ICM soon tr ...

Compared to eutherian mammals, birds retain a primitive form of epiblast development. Molecular studies of the avian epiblast can provide valuable insight for mammalian epiblast research. Here, we introduce several basic techniques in handling epiblast-stage embryos of the chick, ...

Embryonic stem (ES) cells are unique models for investigating early development and cell differentiation. First identified in mouse and later in other mammals, these cells have also been isolated in avian species. Here, using chicken as a model, we describe a set of protocols allowing the isola ...

Whole-mount in situ hybridization (WISH) is a method to visualize gene expression through hybridization of in vitro synthesized riboprobes to cellular mRNAs. WISH has been used in developmental biology for decades and was adapted to many species, especially for model organisms of devel ...

Extracellular matrix can regulate multipotent mesenchymal stromal cells (MMSC) differentiation, with potential applications for tissue engineering. A relief of mineralized bone takes essential effect on MMSC fate. Nevertheless, delicate structure and a hierarchy of nich ...

Marrow stromal cells (MSCs) are relatively rare cells difficult to visualize in marrow biopsies or detect in aspirated marrow. Under specific conditions MSC can be expanded in vitro and the population can give rise to several mesenchymal lineages. “MSC” also refers to mesenchymal stem cells ...

The availability of mesenchymal stem cells (MSCs) or skeletal stem cells (SSCs) is vital to many of the tissue engineering strategies currently being developed for repairing bone and cartilage. One difficulty with using this cell population is that SSCs represent only a small fraction of the ...