细胞技术

The most important tools to investigate the genome of an organism are spontaneous or induced mutations. In mammals, with the exception of humans, the mouse is genetically the most thoroughly analyzed species. Hundreds of different mutant mouse strains are being bred in laboratories and sto ...

Nuclear transfer (NT) is a procedure by which genetically identical individuals can be created. The applications of these NT techniques will be in agriculture, biomedicine, and basic research. Based on the source of donor cells, NT can be classified into embryonic cell NT and somatic cell NT. Som ...

Changes in intracellular calcium concentrations (i) have been shown to regulate a wide variety of developmental processes. In particular, eggs that have completed meiosis or oocytes that are still in the process of undergoing meiotic maturation invariably display some form of a i elevat ...

Fluorescence microscopy has become the method of choice for imaging living specimens, as it offers high signal-to-background and the ability to discriminate between multiple fluorophores. Recently developed techniques, such as confocal (1) or multiphoton imaging (2), permit opt ...

Embryonic stem (ES) cells have the developmental capability to contribute to every cell type of the adult mouse when reintroduced into the blastocyst embryo (1,2). They can be easily manipulated in vitro to produce specific genetic alterations for the study of gene function and the biological ...

Since the early 1980s, embryonic stem (ES) cells have been isolated from the inner cell mass of the preimplantation blastocyst (1–3). They can be maintained in a pluripotent state for indefinite periods of time in the presence of the leukemia inhibitory factor (LIF) or in coculture with mouse embry ...

Primordial germ cells (PGCs) are a small population of unique cells from which all germ cells arise in an organism. In this sense, PGCs can be considered the stem cells of the species. An important characteristic of PGCs is their ability to remain developmentally totipotent,whereas somatic cel ...

cDNA subtractive hybridization is a very powerful method to compare gene expression profiles between two cell or tissue samples of interest. The sample in which differentially expressed transcripts are to be found is usually referred to as “tester,” whereas the reference sample is called ...

RNA interference is a simple, efficient, and highly scalable method for the analysis of gene functions in Caenorhaditis elegans independently of mutants availability (1,2). It is used to study individual genes, gene families, and also for genomewide screenings (3). In a RNAi experiment C. ele ...

The first step in elucidating the function of a particular gene of interest involves defining its expression pattern. One way to do this is by using a reporter gene to tag the gene either in a construct (transgenesis) or in the genome (homologous recombination). Transgenes have been generated using ...

Sea urchins have long been used to study morphogenesis and cell fate specification and are an established model system in developmental biology (1). Most contemporary studies have focused on early development, however, and few molecular genetic studies have examined larval developm ...

Amphibian embryos provide excellent material for understanding the establishment of the vertebrate body plan during early development. Fertilized eggs are readily obtained by hormone-induced spawning, and their developmental rate can be adjusted by ambient temperature re ...

Perhaps the most remarkable recent discovery in developmental biology is that the molecular mechanisms that pattern the animal embryo have been conserved throughout evolution. Many of the genes that specify embryonic pattern were first identified in the fruitfly Drosophila mela ...

For almost two decades, introduction of new genes, transgenic technology, has been the source of many new strains of mice that have become valuable tools in various fields of research. More recently, knockout technology has generated a large number of mutant strains. To maintain all these lines a ...

The development of the cardiovascular system represents one of the earliest critical processes in mammalian development (1). Historically, cardiovascular development has been studied by utilizing in vitro cell- and organ-based systems or by use of avian-based culture models. Even ...

Mammalian embryos grow within the uterus. This inaccessibility makes mammalian embryo investigations more difficult than those using non mammalian embryos. Although attempts to grow mammalian embryos outside of the uterus have appeared in the literature for many decades (see Note ...

Recent technological advances in the generation of recombinant adenovirus vectors have been applied to a wide range of studies in developmental biology. These vectors have allowed analyses of the regulation of expression and function of specific genes as well as evaluation of patterns ...

Human adenovirus is a double-stranded DNA virus with a 36-kb linear genome that contains four early transcription units (E1–E4), active at early (3–12 h) times after infection and five late transcription units (L1–L5) active thereafter and producing the structural genes of the virus. For furt ...

Since Solursh and his coworkers set up the limb bud micromass cultures in 1977 (1), this procedure has become a major model to analyze cellular and molecular event involved in chondrogenesis (2–7). Recently, RCAS retroviral vectors have been developed that can infect chicken embryos in ovo. The ...

Vectors derived from murine retroviruses have been used extensively for gene transfer in both preclinical and clinical studies. Retroviruses are small RNA viruses that replicate through a double-stranded DNA intermediate. The ability of retroviral vectors to integrate effici ...