细胞技术

The production of transgenic laboratory animals has rapidly gained importance as an experimental tool to study the factors that determine the tissue specificity of gene expression, to generate models for human diseases (1,2), as well as to look at the consequences of oncogene expression (3 ...

Introduction of foreign molecules, such as DNA, RNA, or proteins, into living cells is a powerful means to test the biological functions of these molecules. One of the techniques by which foreign molecules can be introduced into living cells is electroporation (for reviews, see refs. 1,2). Compar ...

The ability to transfect mammalian cell lines stably in culture has enabled countless scientific advances to be made in just a few decades. However, not until about 10 years ago did the true potential for the genetic manipulation of cells become a reality, with the ability to alter a mammalian gene pre ...

The introduction of exogenous DNA into mammalian cells can ultimately result in the integration of the transfected DNA into the host genome (1,2). Any genes containing appropriate expression signals will, in most cases, be expressed both prior to and after integration. However, stable exp ...

In many situations, it is important to quantitate accurately expression of a transiently transfected gene. This is most often the case when mechanisms controlling gene expression are under study, and the roles of several possible promoter and enhancer elements and their interacting fa ...

The efficient production of proteins via expression of recombinant genes in cells other than those that naturally produce the protein has been critical to the study and the therapeutic utility of a wide variety of proteins. E. coli expression systems are useful for many proteins. However, large ...

The ability to introduce DNA into embryos for the purpose of obtaining stable transformants is an important part of the technology that supports the exquisite genetic and molecular studies in Drosophila. As this ability to stably transform Drosophila embryos was developed, it became obv ...

Fish are the largest group of vertebrates. They have diverse features to adapt to a wide variety of environments, which make them excellent models for studies in various areas, including molecular and cellular biology (1,2). Small freshwater species, such as medaka (Oryzias latipes) and zebr ...

High-intensity, pulsed electrical shock is deliberately applied to the heart in the clinical setting for the treatment of cardiac arrhythmia, e.g., during defibrillation. However, overdoses of electrical shock can result in toxic instead of therapeutic effects (1). Successful defi ...

Interest continues to grow rapidly in clinical gene therapy. Although many protocols approved by the US NIH Recombinant Advisory Committee involve the genetic marking of cells to determine, for example, the source of relapse after autologous bone marrow transplantation for certain m ...

Proteins can be introduced into cells in several ways, including permeabilization by Sendai virus, trypsinization, osmotic shock, microinjection, electroporation, or after transfection of cells with expression vectors containing the gene(s) of interest. Unlike most of these o ...

Specific cytotoxic T-lymphocytes (CTL) are important in the protection against viral and parasitic infections (1,2), as well as monitoring and even eradicating tumor cells (3–5). CTL normally recognize endogenously processed peptide antigens (Ag) complexed to MHC class I molecules ...

The introduction into mammalian cells of antibodies with specificities for endogenous cellular factors permits the functional assessment of such factors in the context of living cells. Antibodies have been successfully introduced into several cell types by various methods, inc ...

Electroporation has been used for the introduction of DNA (1–4), proteins (5–7), and various nonpermeant drugs and metabolites into cultured mammalian cells (8,9), as described in this and other volumes (10,11). Most electroporation techniques for adherent cells involve the delivery of t ...

Chinese hamster ovary (CHO) cells possess many advantages that make them among the most widely used and highly characterized mammalian cell lines. They are easy to culture, they grow rapidly without density dependence, and they do not have stringent culture medium or high serum growth requir ...

Electroporation has become a useful tool for the introduction of biomolecules to a variety of cell types since the method was first described (1,2). This technique has also been used for important functional studies in the different GH cell strains: rat pituitary tumor cells that produce prola ...

Electrofusion has developed into an extremely efficient method for the fusion of mammalian cells. Cell-cell fusion has become an important tool for the study of cell biology, molecular biology, and bioproduction of important biological substances. Although other fusion methodol ...

The methods of induced cell fusion are very useful procedures in reproductive and developmental biology. They are used to answer basic questions associated with cell-cycle regulation in mammalian oocytes and embryos (1,2), to produce tetraploid embryos (3), and of particular interest, ...

Epstein Barr virus (EBV) can be used to transform human B-lymphocytes to derive populations of cells secreting specific antibodies of interest. Isolating monoclonal or stable populations of these cells, however, has proven very difficult (1). In our laboratory, we have developed methods ...

In early work to produce identical animals, such as twins, triplets, or quadruplets, blastomeres of two- to four-cell stage sheep embryos were separated (1), and each blastomere was sealed in an empty zona pellucida, embedded in agar gel, and temporarily cultured in vivo in rabbit or sheep oviducts u ...