细胞技术

Cellular senescence processes affecting tissue resident stem cells are considered, at present, an hallmark of both aging and age-related pathologies. Therefore it is mandatory to address this problem with adequate techniques that could highlight the molecular alterations asso ...

We describe protocols for the isolation of satellite cells from human muscle biopsies, for the in vitro culture of proliferating and differentiating myoblasts, and for the preparation of cell samples suitable for morphological and cytochemical analyses at light and electron micros ...

Our lab developed and optimized a method, known as the modified pre-plate technique, to isolate stem/progenitor cells from skeletal muscle. This method separates different populations of myogenic cells based on their propensity to adhere to a collagen I-coated surface. Based on their su ...

Many chronic liver diseases are life-threatening. When the liver loses the ability to repair itself the only treatment currently available is liver transplant. However, there are not enough donors to treat all the patients. This requires the search of alternative therapies utilizing st ...

Hematopoietic stem cells (HSCs) are capable of self-renewal and multi-lineage reconstitution of hematopoiesis in irradiated transplant recipient mice. As such, bone marrow transplantation (BMT) is a major assay commonly used to examine murine HSC activity. BMT traditionally in ...

Hematopoietic stem cells (HSCs) have the ability to self-renew and replenish the blood and immune system for the life span of an individual. An age-associated decline in HSC function is responsible for the decreased immune function and increased incidence of myeloid diseases and anemia in the ...

The use of flow cytometry has been critical in establishing methods to isolate and characterize hematopoietic stem cells (HSCs) and their progenitors. For more than 30 years, researchers have been uncovering novel markers that when used in combination significantly enhance the purif ...

Quantitative fluorescence in situ hybridization (Q-FISH) is a complex technique for the quantitative evaluation of telomere length on cell preparations or on human tissues. The samples are stained with a fluorescent peptide nucleic acid (PNA) probe against the telomere oligonucle ...

Epigenetic regulation is one of the major players in gene expression control. Histone modifications including acetylation or methylation are epigenetic mechanisms known to regulate senescence associated genes expression during adult stem cell aging. Chromatin immunopre ...

Long-term culture of mesenchymal stem cells (MSC) has major impact on cellular characteristics and differentiation potential. Numerous clinical trials raise high hopes in regenerative medicine and this necessitates reliable quality control of the cellular products—also wi ...

In vivo transplantation of putative populations of hematopoietic stem cells (HSC) and assessment of their engraftment is considered the golden standard to assess their quality and degree of stemness. Transplantation is usually carried out by intravenous injection in murine models ...

RNA interference (RNAi) is a powerful gene-knockdown technology that has been applied for functional genetic loss-of-function studies in many model eukaryotic systems, including embryonic stem cells (ESCs). Application of RNAi in ESCs allows for dissection of mechanisms by which E ...

Adenosine is an important modulator of metabolic activity with powerful tissue- and cell-protective functions. Adenosine kinase (ADK), the major adenosine-regulating enzyme, is critical to adapt its intra- and extra-cellular levels in response to environmental changes. Lenti ...

Quantitative real-time PCR (qRT-PCR) is a reliable method to determine and monitor microRNA (miRNA) expression profiles in different cells, tissues, and organisms. Although there are several different strategies for performing qRT-PCR to determine miRNA expression, all of them ha ...

A more complete understanding of the biology of adult stem cells could yield important insights toward devising effective cell-based regenerative therapies to treat disease. The germline stem cells (GSCs) in the fruit fly Drosophila melanogaster are an excellent in vivo model for the st ...

MicroRNAs are key regulators of gene expression in diverse biological processes and their importance in embryonic stem cells is indisputable. New ‘next-generation’ technologies such as Illumina massively parallel sequencing offer vast improvements, in both scale and sensiti ...

RNA interference (RNAi) technology is one of the main means in the study of stem cell differentiation. This study describes Rho-GDIγ function during the differentiation of neural stem cells by using RNAi. Rho-GDIγ belongs to the Rho-GDI protein family, which is expressed at high level through ...

Techniques for controlling the expression of a specific gene in embryonic stem cells are effective and important for clarifying the functions of the gene. Regarding differentiation of cells into nervous system components, these techniques would play key roles in elucidating, not only ...

RNA interference is widely used for loss-of-function studies in mammalian cells. As an alternative to the transfection of small RNAs, plasmid vectors have been developed to express short hairpin RNAs (shRNAs). We engineered the pHYPER shRNA vector, which is based on a 2.5-kb mouse genomic frag ...

The following protocols provide a rapid approach for establishing good working conditions for transfecting siRNAs for specific gene knockdown. By first using microscopy to evaluate efficient transfection of an inexpensive, fluorescent oligonucleotide, the researcher can ...