细胞功能测定

Human embryonic stem cell lines are usually derived from human embryos that have become excess to clinical needs in assisted reproduction programs, whether because the couple in question has completed their family or because the embryo was found to be clinically unsuitable for transfer d ...

The majority of human embryonic stem cell (hESC) lines have been derived and grown using mouse or human feeder cells, or using Matrigel�, an animal derivative rich in extracellular matrix (ECM) proteins. However, reliance on feeder layers and animal products limits the manipulation and clin ...

Human embryonic stem cells (hESCs) have the capacity to self-renew and to differentiate into all components of the embryonic germ layers (ectoderm, mesoderm, endoderm) and subsequently all cell types that comprise human tissues. HESCs can potentially provide an extraordinary source ...

We describe in this chapter the development of a xenofree molecularly defined medium, SBX, associated with xenofree matrices, to maintain human embryonic stem cell (hESC) pluripotency as determined by phenotypic, functional and TLDA studies. This simple, inexpensive, and more physi ...

Human embryonic stem cells (hESCs) are pluripotent stem cells derived from the inner cell mass of human blastocysts. hESCs have become a great asset to studying human diseases and genetic functions of healthy organisms. The rate at which hESCs are being used in laboratories is exponentially i ...

Human embryonic stem cells (hESC) involve long-term cultures that must remain undifferentiated. The real-time PCR (RT-PCR) technique allows the relative quantification of target genes, including undifferentiation and differentiation markers when referred to a housekeep ...

The routine culture and expansion of human embryonic stem (hES) cells has been and is still posing a challenge to researchers wishing to take advantage of the cells' unique potential. In contrast to mouse embryonic stem cells, hES cells usually have to be expanded by tedious mechanical microdiss ...

Progress from our present understanding of the mechanisms behind mitosis has been compromised by the fact that model systems that were ideal for molecular and genetic studies (such as yeasts, C. elegans, or Drosophila) were not suitable for intracellular micromanipulation. Unfortun ...

During cell division microtubules of the mitotic spindle segregate the duplicated chromosomes into the two daughter cells. Chromosome–microtubule attachment is mediated by kinetochores, multiprotein complexes assembled on specialized regions of the DNA. Kinetochores m ...

Tissue microarrays enable the rapid histological localization of gene expression in hundreds of archival samples by in situ hybridization. However, the scoring of tissue microarray data may be influenced by intra- and inter-observer variations, and categorizing continuous var ...

In situ hybridization is a powerful technique for examining changes in mRNA levels. Its primary advantage over the Northern blot and reverse transcription polymerase chain reaction is the ability to localize specific species of mRNA to a particular cell population in a heterocellular s ...

This chapter explores the combination of a nonradioactive in situ hybridization technique to detect mRNA with an imunnohistochemical labeling technique for use in formalin- fixed, paraffin-embedded, or frozen tissue sections. This technique allows the combination of detecti ...

Cell death by apoptosis is now recognized widely as an important constituent of cell turnover and disease pathology. Characterized by the cleavage of DNA into oligonucleosome-sized DNA fragments, end-labeling of fragmented DNA often is used as an in situ histological marker of apoptosi ...

Monoclonal antibodies to proliferation associated antigens have long been used to histologically localize mitogenesis. However, techniques that distinguish cells in the synthetic or S phase have tended to rely on the in vivo incorporation of tritiated thymidine or thymidine anal ...

In situ hybridization is a basic method in modern plant cell and molecular biology. It is used to locate the chromosomal position of genomic DNA sequences. It is able to determine the patterns of gene transcription in mature tissues and during development. In situ hybridization, in combination w ...

The potential of cell and gene therapy has generated extensive interest over the past several years. More recently, identification of stem cells of various types, especially embryonic stem cells, reinforced this interest. Systematic studies are now being launched to define the biology ...

In this chapter, we describe a simple and relatively rapid technique for detecting lowabundance slug mRNA in cultured cells. The procedure uses nonradioactive digoxigeninlabeled RNA probes that are more sensitive than deoxyribonucleic probes and simpler to detect than radioact ...

A method is described for in situ hybridization of riboprobes to free-floating brain sections. Brain sections are hybridized and processed free-floating in buffer, i.e., without attachment to a support such as a slide. To withstand the extra wear compared with sections processed on-slide, t ...

This chapter describes a pre-embedding in situ hybridization method utilizing an immunogold-silver intensification step to identify P2Y2 receptor mRNA transcripts in the adult rat cerebellum. The method was applied for ultrastructural (electron microscopic) examinatio ...

The term in situ hybridization (ISH) refers to all methods allowing the detection of specific DNA (gene loci) or RNA (gene expression products) sequences, using molecular hybridization (base pairing) of labeled nucleic acid probes to target molecules within “intact” cell populations in ...