细胞技术

Directed differentiation of human embryonic stem cells (hESCs) to a functional cell type, including neurons, is the foundation for application of hESCs. We describe here a reproducible, chemically defined protocol that allows directed differentiation of hESCs to nearly pure neuro ...

Traditional methods of studying the differentiation of human embryonic stem cells (hESCs) include generation of embryoid bodies, induced differentiation in vitro, and transplantation to immune-deficient mice. The chick embryo is a well-studied and accessible experimental s ...

The culture and critical assessment of early human embryos during the first week of human development are reviewed for the derivation of ESC. Both normal and abnormal features are assessed by phase contrast microscopy of whole embryos and in serial sections of fixed material by light and elect ...

One of the great advantages of embryonic stem (ES) cells over other cell types is their accessibility to genetic manipulation. They can easily undergo genetic modifications while remaining pluripotent and can be selectively propagated, allowing the clonal expansion of genetically ...

Generic methods for genetic manipulation of human embryonic stem cells (hESCs) are important for both present research and future commercial applications. To date, differences in cell derivation and culture have required independent optimization of transfection and transdu ...

The recent discovery of genomic reprogramming of human somatic cells to an embryonic stem (ES) cell-like pluripotent state provides a unique opportunity for stem cell research. The reprogrammed cells, named as induced pluripotent stem (iPS) cells, possess many of the properties of ES cells ...

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 ...