细胞技术

Human bone marrow (BM) contains a population of non-hematopoietic stem cells also termed stromal cells, mesenchymal cells or multipotent mesenchymal stromal cells (MSCs). These cells have unique stem cell-like properties including their ability to self-renew, differentiate in ...

Mycoplasma contamination of cell lines is one of the major problems in cell culture technology. The �specific, sensitive, and reliable detection of mycoplasma contamination is an important part of mycoplasma control and should be an established method in every cell culture laboratory. ...

We describe a method to derive cell lines and clones from cells of the murine midgestation aorta-gonads-mesonephros (AGM) microenvironment. We start from subdissected AGM regions in “explant” or “single cell suspension” type cultures from embryos transgenic for tsA58, a temperature ...

Leukocyte recruitment from the vasculature occurs under conditions of haemodynamic shear stress. The parallel plate flow chamber apparatus is an in vitro system that is widely used to study leukocyte recruitment under shear conditions. The flow chamber is a versatile tool for examining ...

Hematopoietic stem cells (HSCs) present in small numbers in adult bone marrow (BM), peripheral blood (PB) and umbilical cord blood (CB) produce a heterogeneous pool of progenitors that can be detected in vitro using colony forming cell (CFC) assays. Hematopoietic progenitor cells prolif ...

The long-term culture-initiating cell (LTC-IC) assay is a well-established in vitro assay used to enumerate primitive mouse hematopoietic stem cells (HSCs) and relies on the two cardinal functions of HSCs: ability to self-renew and differentiation capacity. LTC-ICs present in minima ...

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 ...