细胞技术

The majority of lymphomas originate from B cells at the germinal center stage or beyond. Preferential selection of B cell clones by a limited set of antigens has been suggested to drive lymphoma development. However, little is known about the specificity of the antibodies expressed by lymphoma ...

The assessment of the presence of clonal lymphoproliferations via polymerase chain reaction (PCR)-based analysis of rearranged immunoglobulin (Ig) or T-cell receptor (TCR) genes is a valuable technique in the diagnosis of suspect lymphoproliferative disorders. Furthermore ...

Normal and malignant B cells carry rearranged immunoglobulin (Ig) variable region genes, which due to their practically limitless diversity represent ideal clonal markers for these cells. We describe here an approach to isolate single cells from frozen tissue sections by microdiss ...

Multiparametric flow cytometry is a powerful diagnostic tool that permits rapid assessment of cellular antigen expression to quickly provide immunophenotypic information suitable for disease classification. This chapter describes a general approach for the identific ...

Immunoglobulin (Ig) gene remodeling by V(D)J recombination plays a central role in the generation of normal B cells, and somatic hypermutation and class switching of Ig genes are key processes during antigen-driven B cell differentiation. However, errors of these processes are involved ...

High-throughput mRNA sequencing (RNA-seq) uses massively parallel sequencing to allow an unbiased analysis of both genome-wide transcription levels and mutation status of a tumor. In the RNA-seq method, complementary DNA (cDNA) is used to generate short sequence reads by immobilizi ...

The herpesviruses Epstein-Barr virus (EBV) and human herpesvirus 8 and the retrovirus human T-cell leukemia virus type 1 are directly implicated in the pathogenesis of lymphoma and leukemia in man. EBV is associated with an expanding spectrum of lymphomas and it would appear likely that addi ...

MicroRNAs (miRNAs) play important roles in development, differentiation, homeostasis, and also in diseases such as lymphoma. This chapter describes methods to study the role of miRNAs in lymphoma. First, we describe a multiplex RT reaction followed by qPCR that can be used to determine diff ...

RNA interference (RNAi) is a conserved posttranscriptional gene silencing mechanism that has recently emerged as a breakthrough genetic tool in functional genomics and drug target discovery. An increasing number of studies applying RNAi in high-throughput screens have begun to u ...

Fluorescence In Situ Hybridization (FISH) is a powerful and robust technique allowing the visualization of target sequences like genes in interphase nuclei. It is widely used in routine diagnostics to identify cancer specific aberrations including lymphoma associated translo ...

Through the genome-wide characterization of a cell type’s transcriptome, gene expression profile analysis provides a potent tool for analyzing the pathogenesis of lymphomas and has had a major impact on the understanding of lymphoid neoplasia. The analysis of gene expression patter ...

Normal DNA methylation is an epigenetic modification required for proper development. Aberrant DNA methylation, in contrast, is frequently observed in many different malignancies including leukemias and lymphomas. Global DNA methylation profiling addresses the methyla ...

Minimal residual disease (MRD) diagnostics is of high clinical relevance in patients with indolent B-cell Non-Hodgkin lymphomas (B-NHL) and serves as a surrogate parameter to evaluate treatment effectiveness and long-term prognosis. MRD diagnostics performed by real-time quan ...

Cancer genes are often deregulated by genomic rearrangements. Accordingly, analysis of the participant chromosomes responsible now occupies a key role in characterizing and identifying cancer cell lines. Cytogenetics may also be used to study the nature and extent of chromosome br ...

The main prerequisition for any research, development, or production programs involving cell lines is whether a cell line is authentic or not. Microsatellites in the human genome harboring short tandem repeat (STR) DNA markers allow the identification of individual cell lines at the DNA le ...

Studies of the same cell lines by different laboratories are common in the literature and often show different results with the same methodology. Use of best cell culture practices is essential to ensure consistent and reproducible results. Assay outcomes are easily influenced by many fac ...

The successful storage of cell lines depends upon many factors, including the condition of the cells to be frozen and the experience of the operator. Attempting to freeze down unhealthy, contaminated or poorly labelled cells can have huge implications for a research laboratory. This chapter ...

Three experimental systems based on mouse models are currently used to study breast cancer: transgenic mice, carcinogen-induced models, and xenografts of breast cancers. Each of these models has advantages and limitations. This chapter focuses on xenotransplantation of breast ca ...

Maintenance of a mitotically inactive feeder layer which is able to provide extracellular matrix and growth factors can be critical in establishing and maintaining primary tumor cells. How feeder cells are handled and processed is crucial for providing trouble-free support for prima ...

Drug design is an iterative process requiring cycles of compound synthesis and testing, with each successive synthesis phase yielding molecules predicted to have improved characteristics over the previous set of compounds. In the field of cancer drug discovery, a key early-stage elem ...