细胞功能测定

Pluripotent stem cells (PSCs) derived from parthenogenetically activated human oocytes demonstrate the typical characteristics displayed by human embryonic stem cells (hESCs) including infinite division and in vitro and in vivo differentiation into cells of all germ lineag ...

Human embryonic stem cells (hESCs) are self-renewing, pluripotent cells that serve as a valuable research tool and hold promise for use in regenerative medicine. Most hESC lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in ex ...

The ability to cryopreserve and successfully recover cell lines has been critical to the conservation of all cell lines, especially the preservation of pristine early-stage cultures and the preparation of well-characterized cell banks. Indeed, the systematic storage and establi ...

This chapter describes some of the major issues to be considered when setting up a laboratory for the culture of human pluripotent stem cells (hPSCs). The process of establishing a hPSC laboratory can be divided into two equally important parts. One is completely administrative and includes d ...

Direct MALDI-TOF mass spectrometric peptide profiling is increasingly used to analyze the peptide complement in the nervous system of a variety of invertebrate animals, from leech to Aplysia and many arthropod species, especially insects and crustaceans. Proper sample preparati ...

With the entire genome sequence of several animals now available, it is becoming possible to identify in silico all putative peptides and their precursors in an organism. In this chapter we describe a searching algorithm that can be used to scan the genome for predicted proteins with the structur ...

Polypeptide and protein arrays enable high-throughput screening capabilities for studying molecular interactions and profiling of biomarkers, and provide a powerful functional screening tool for peptidomics. To overcome the limitations of conventional arraying meth ...

The transparent soil nematode Caenorhabditis elegans can be considered an important model organism due to its ease of cultivation, suitability for high-throughput genetic screens, and extremely well-defined anatomy. C. elegans contains exactly 959 cells that are ordered in defin ...

Host–pathogen interactions are complex competitions during which both the host and the pathogen adapt rapidly to each other in order for one or the other to survive. Salmonella enterica serovar Typhimurium is a pathogen with a broad host range that causes a typhoid fever-like disease in mice and ...

Recent years have seen great advances in mass spectrometry and proteomics, the science dealing with the analysis of proteins, their structure and function. A branch of proteomics dealing with naturally occurring peptides is often referred to as peptidomics. Direct analysis of peptides ...

A key challenge in clinics is the identification of sensitive and specific biomarkers for early detection, prognostic evaluation, and surveillance of disease. A biomarker is defined as a biological substance that can be used to specifically detect a disease, measure its progression, or t ...

An emerging way to study neuropsychiatric or neurodegenerative diseases is by performing proteomic analyses of brain tissues. Here, we describe methods used to isolate and identify the proteins associated with a sample of interest, such as the synapse, as well as to compare the levels of prote ...

Due to the complexity of the mammalian central nervous system, neuropeptidomic studies in mammals often yield very complicated mass spectra that make data analysis difficult. Careful sample preparation and extraction protocols must be employed in order to minimize spectral compl ...

Here we report our approach to the peptidomic analysis of mouse liver. We use ultrafiltration for peptide prefractionation, which is followed by size exclusion chromatography. The low molecular weight peptides (MW below ~3kDa) are analysed directly by nanoLC-MS/MS, and the higher mole ...

Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in ...

A large number of diverse antimicrobial peptides have been found in amphibian skins, and many more remain to be identified. It is sufficiently easy to obtain amounts of gland secretions sufficient for both identification and functional testing of the bioactive peptides. We describe here a s ...

Antimicrobial peptides (AMPs) play an important role in the host’s innate defence system in many organisms. Amphibian skin is expected to be a particularly rich source of novel AMPs. In amphibians, AMPs are produced from precursor proteins via specific cleavage by processing enzymes. While ...

Skin secretions from anurans (frogs and toads), particularly those species belonging to the Hylidae and Ranidae families, are a rich source of biologically active peptides. Cytolytic peptides with broad-spectrum antimicrobial activities and highly variable amino acid sequen ...

Today, commercially available mass spectrometers increasingly meet all the demands of the proteomics community including high throughput, high sensitivity, and significant fragmentation capability for sequence determinations. Therefore, proper sample preparati ...

Direct MALDI-TOF mass spectrometric peptide profiling is increasingly used to analyze the peptide complement in the nervous system of a variety of invertebrate animals from leech to Aplysia and many arthropod species, especially insects and crustaceans. Here, we describe a protocol f ...