细胞功能测定

The long-term culture initiating cell (LTC-IC) assay, founded on the bone marrow long-term culture (LTC) system, measures primitive hematopoietic stem cells (termed LTC-IC) based on their capacity to produce myeloid progeny for at least 5 weeks. Adaptations of the LTC system including the u ...

Macrophages play a key role in the innate immune response and help to direct the acquired immune response. Early in the innate immune response, they produce reactive oxygen species and pro-inflammatory cytokines and chemokines to drive inflammation and are referred to as “classically ac ...

Megakaryocytes (MK) are hematopoietic cells present in the bone marrow that are responsible for the production and release of platelets in the circulation. Given their very low frequency (

The cancer stem cell hypothesis is an appealing concept to account for intratumoral heterogeneity and the observation that systemic metastasis and treatment failure are often associated with the survival of a small number of cancer cells. Whilst in vivo evidence forms the foundation of t ...

Hoechst side population (SP) analysis remains a critical technique for identifying stem cell and progenitor populations in hematopoietic and non-hematopoietic tissues, as well as potential cancer stem cells. More recently, DyeCycle Violet (DCV), a DNA binding dye structurally si ...

The side population (SP) is a subpopulation of mouse bone marrow cells highly enriched for hematopoietic stem cell activity. The SP is identified using flow cytometry as a minor population that efficiently effluxes the DNA-binding dye Hoechst 33342 relative to the rest of the bone marrow. Phen ...

Ascidians (marine invertebrates: urochordates) are thought to be the closest sister groups of vertebrates. They are particularly attractive models because of their non-duplicated genome and the fast and synchronous development of large populations of eggs into simple tadpoles m ...

One of the most straightforward approaches to making novel biological discoveries is the forward genetic screen. The time is ripe for forward genetic screens in the mouse since the mouse genome is sequenced, but the function of many of the genes remains unknown. Today, with careful planning, such ...

Establishment of methods to inactivate genes by homologous recombination in embryonic stem (ES) cells has provided great advantages to the field of mouse genetics. Using this technology, a number of null mutant mice, so-called knock-out mice, have been generated. The gene-targeting tec ...

Nonhuman primates (NHP) are the closest animal species to humans and have been widely used for studying human reproductive physiology. Assisted reproductive technology (ART) in Old World NHPs provides great opportunity for studying fertilization, embryo development, embryon ...

Analysis of gene expression patterns is central to the study of embryonic development. This chapter details methods for detecting gene expression in whole mouse embryos and in tissue sections. The most commonly used methods available in mouse are described and include mRNA in situ hybridi ...

Ascidians, such as Ciona, are invertebrate chordates with simple embryonic body plans and small, relatively non-redundant genomes. Ciona genetics is in its infancy compared to many other model systems, but it provides a powerful method for studying this important vertebrate outgroup. ...

Reptiles (lizards, snakes, turtles and crocodylians) are becoming increasing popular as models for developmental investigations. In this review the leopard gecko, Eublepharis macularius, is presented as a reptilian model for embryonic studies. We provide details of husbandry, b ...

The invertebrate chordate amphioxus (Branchiostoma), which is the most basal living chordate, has become an accepted model for the vertebrate ancestor in studies of development and evolution. Amphioxus resembles vertebrates in regard to morphology, developmental gene expres ...

The developing marsupial is an ideal animal for use in biomedical research. Marsupials are mammals that have been separated from eutherian mammals for over 130–140 million years. They all deliver altricial young that complete their growth and development after birth usually in a pouch, but n ...

TILLING (Targeting Induced Local Lesions IN Genomes) is a popular reverse genetic approach that has been successfully applied in several genetic model organisms such as zebrafish, rat, Drosophila, Arabidopsis, or medaka. In contrast to classical targeted knockout technologies th ...

Recent debates about model organisms echo far into the past; taking a longer view adds perspective to present concerns. The major approaches in the history of research on vertebrate embryos have tended to exploit different species, though there are long-term continuities too. Early ninet ...

Animal models, including the zebrafish, without a reliable embryonic stem cell system are not easily amenable to targeted mutagenesis for studying gene function. Three recent publications have shown that zinc finger nucleases (ZFNs) have circumvented this shortcoming in zebrafi ...

This chapter describes a detailed protocol for deriving mouse embryonic stem (ES) cell lines from blastocysts that are produced either by fertilization or by somatic cell nuclear transfer (NT or cloning). Rather than function as a scientific communication with the experts in this field, th ...

For two decades, my lab has been studying development using two vertebrate animals, the frog Xenopus and the zebrafish, Danio. This has been both productive and challenging. The initial rationale for the choice was to compare the same process in two species, as a means to find commonalities that may ca ...