细胞技术

Animal models have added significantly to our understanding of the mechanism(s) of hematopoietic stem and progenitor cell (HSPC) mobilization. Such models suggest that changes in the interaction between the HSPC and the hematopoietic microenvironmental ‘niche’ (cellular and e ...

Understanding mechanisms responsible for engraftment of hematopoietic stem cells (HSC) is important to achieve successful HSC transplantation. Homing of HSC to the bone marrow niche is believed to be a crucial step for engraftment. However, the molecular mechanisms that regulate HSC ...

The balance between the quiescent hematopoietic stem cell (HSC) and the highly proliferative hematopoietic progenitor compartments maintains homeostasis in the hematopoietic system. Therefore, the entry of HSCs into the cell cycle and the rate of proliferation of hematopoiet ...

Analysis of apoptosis can be used to assess aging and survival in the hematopoietic stem cell (HSC) compartment in the context of disease, therapeutic manipulation, or genetic mutation. Two different methods to assess the frequency of apoptosis in the HSC compartment are presented. The fir ...

Various assays exist that measure the function of hematopoietic stemcells (HSCs). In this chapter, in vitro assays are described that measure the frequency of progenitors (colony-forming unit in culture; CFU-C), stem cells (long-term culture-initiating cell; LTC-IC), or both (cobble ...

In recent years, the field of stem cells has become one of the most rapidly growing areas in biological and medical sciences. Embryonic stem (ES) cells differentiate efficiently in vitro and give rise to many different somatic cell types. The ability to generate a wide spectrum of differentiated c ...

Traditionally, the study of human T cell development has relied on the availability of human and mouse thymic tissue. In this chapter, we outline a simple in vitro protocol for generating large numbers of human T-lineage cells from umbilical cord blood (CB)- derived hematopoietic stem cells (HS ...

The successful isolation and characterization of human embryonic stem cells (hESCs) provides a powerful tool to study the cellular and genetic mechanisms that mediate cell-fate decisions toward distinct developmental lineages. hESC-derived cells may also be suitable for novel c ...

In traditional bone marrow transplantation, the majority of peripherally introduced stem cells are trapped in peripheral organs, such as the lung and liver. The frequency of cells homed in bone marrow by such method is extremely low. This circumstance adds difficulty to the research of hemat ...

The “side population” (SP) phenotype is a manifestation of primitive cells’ ability to efficiently efflux the fluorescent DNA-staining dye Hoechst 33342 and can be used as the basis by which to isolate these cells using flow cytometry. In the bone marrow (BM), the SP defines a cell subset with a highly h ...

Complementing mutant embryos or embryonic stem cells with normal cells in embryonic chimeras is a valuable tool for investigating phenotypes. Chimera approaches provide a method to examine the phenotype of mutant cells, including hematopoiesis, in mutants with early embryonic le ...

Hematopoietic stem cell transplantation has traditionally been used to reconstitute blood cell lineages that had formed abnormally because of genetic mutations, or that had been eradicated to treat a disease such as leukemia. However, in recent years, much attention has been paid to the new ...

Hematopoietic stem cells (HSC) are inherently rare cell types that cannot be obtained in sufficient amounts for classical biochemical characterization. To facilitate functional studies of murine HSC and hematopoietic development, the technique of retroviral-mediated gene ...

The birth of viable offspring from somatic cell nuclear transfer (SCNT) in mammals caused a major re-examination of the understanding of the commitment of cells to specific tissue lineages during differentiation. The questions of whether cells undergo dedifferentiation or transd ...

This chapter describes a procedure for isolation and analysis of fractions enriched in plasma membranes from minute amounts of tissue. It consists of a method for extraction and fractionation of membranes and a method for enzymatic digestion of membrane proteins without use of detergen ...

Sequential detergent extraction of proteins from eukaryotic cells has been used to increase proteome coverage of 2D-PAGE. We have adapted sequential detergent extraction for use with the high-throughput non-electrophoretic proteomics method of liquid chromatography and el ...

Mitochondria are essential organelles in cellular metabolism. These organelles are bounded by two membranes, the outer and inner membrane. Especially the inner membrane comprises a high content of proteins, for example, the protein complexes of the respiratory chain. High-resolu ...

Identification and characterization of membrane proteins is of increasing importance in modern proteomic studies. It is of central interest to have access to methods that combine efficient solubilization with enrichment of proteins and intact protein complexes. Separation me ...

Generally, a combination of two or more chromatographic and/or electrophoretic methods is conducted to separate membrane protein complexes. Here we describe how thylakoid membrane protein complexes from the photosynthetic apparatus can be successfully separated by two main s ...

Pancreatic zymogen granules (ZG) are specialized for digestive enzyme storage and regulated secretion in the exocrine pancreas and are a classical model for studying secretory granule function. To understand the function of this organelle, we have conducted a proteomic study to iden ...