Exosome, Apoptosis and Stem-cell therapy and Clinical Diagnosis with Flow Cytometry
https://en.wikipedia.org/wiki/Apoptosis
https://en.wikipedia.org/wiki/Exosome_(vesicle)
https://en.wikipedia.org/wiki/Stem-cell_therapy
https://en.wikipedia.org/wiki/Flow_cytometry

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Exosomes are cell-derived vesicles that are present in many, and perhaps all, eukaryotic fluids including blood, urine, cerebrospinal fluid and cultured medium of cell cultures.[1][2][3] The diameter of exosomes is between 30 and 100 nm, which is larger than low-density lipoproteins (LDL) but smaller than microvesicles and much smaller than cells.[1] Exosomes are either released from the cell when multivesicular bodies fuse with the plasma membrane or released directly from the plasma membrane.[4] Evidence is accumulating that exosomes have specialized functions and play a key role in processes such as coagulation, intercellular signalling, and waste management.[1] Consequently, there is a growing interest in the clinical applications of exosomes. Exosomes can potentially be used to inform prognosis, for therapy, and as biomarkers for health and disease.[5]. https://en.wikipedia.org/wiki/Exosome_(vesicle)

Bioinformatics analysis
Exosomes contain RNA, proteins, lipids and metabolites that is reflective of the cell type of origin. As exosomes contain numerous proteins, RNA and lipids, large scale analysis including proteomics and transcriptomics is often performed. Currently, to analyse these data, non-commercial tools such as FunRich^[52] can be used to identify over-represented groups of molecules. With the advent of Next generation sequencing technologies, the research on exosomes have been accelerated in not only cancer but various diseases. Recently, bioinformatics based analysis of RNA-Seq data of exosomes extracted from Trypanosoma cruzi has showed the association of these extracellular vesicles with various important gene products that strengthens the probability of finding biomarkers for Chagas disease.^{[53][54], ..}
https://en.wikipedia.org/wiki/Exosome_(vesicle)#Bioinformatics_analysis

Therapeutics and carriers of drugs
Increasingly, exosomes are being recognized as potential therapeutics as they have the ability to elicit potent cellular responses in vitro and in vivo.^[55][56][57] Exosomes mediate regenerative outcomes in injury and disease that recapitulate observed bioactivity of stem cell populations.^[58]Mesenchymal stem cell exosomes were found to activate several signaling pathways important in wound healing (Akt, ERK, and STAT3) and bone fracture repair.^[59][60] They induce the expression of a number of growth factors (hepatocyte growth factor (HGF), insulin-like growth factor-1(IGF1), nerve growth factor (NGF), and stromal-derived growth factor-1 (SDF1)).^{[61], ..}
https://en.wikipedia.org/wiki/Exosome_(vesicle)#Therapeutics_and_carriers_of_drugs

In contrast to necrosis, which is a form of traumatic cell death that results from acute cellular injury, Apoptosis is a highly regulated and controlled process that confers advantages during an organism's lifecycle. For example, the separation of fingers and toes in a developing human embryo occurs because cells between the digits undergo apoptosis. Unlike necrosis, apoptosis produces cell fragments called apoptotic bodies that phagocytic cells are able to engulf and remove before the contents of the cell can spill out onto surrounding cells and cause damage to them.[5]. .. https://en.wikipedia.org/wiki/Apoptosis

Implication in disease :    Defective pathways, Dysregulation of p53, Inhibition, HeLa cell, Hyperactive apoptosis, HIV progression, Viral infection, Stem-cell therapy, Bone marrow transplant, ..

Stem-cell therapy is the use of stem cells to treat or prevent a disease or condition.[1] Bone marrow transplant is the most widely used stem-cell therapy, but some therapies derived from umbilical cord blood are also in use. Research is underway to develop various sources for stem cells, as well as to apply stem-cell treatments for neurodegenerative diseases and conditions such as diabetes and heart disease, among others. Applications : Neurodegeneration, Brain and spinal cord injury, Heart, Blood-cell formation, Regrowing teeth Cochlear hair cell regrowth, Blindness and vision impairment, Pancreatic beta cells, Orthopedics Wound healing, Infertility, HIV/AIDS, Regenerative treatment models, Drug discovery and biomedical research, .. https://en.wikipedia.org/wiki/Stem-cell_therapy

Veterinary medicine Research has been conducted on horses, dogs, and cats can benefit the development of stem cell treatments in veterinary medicine and can target a wide range of injuries and diseases such as myocardial infarction, stroke, tendon and ligament damage, osteoarthritis, osteochondrosisand muscular dystrophy both in large animals, as well as humans.[74][75][76][77] While investigation of cell-based therapeutics generally reflects human medical needs, the high degree of frequency and severity of certain injuries in racehorses has put veterinary medicine at the forefront of this novel regenerative approach.[78] Companion animals can serve as clinically relevant models that closely mimic human disease. .. https://en.wikipedia.org/wiki/Stem-cell_therapy#Veterinary_medicine

Flow cytometry is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles.[1][2][3][4] A sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam and the light scattered is characteristic to the cells and their components. Cells are often labeled with fluorescent markers so that light is first absorbed and then emitted in a band of wavelengths. Tens of thousands of cells can be quickly examined and the data gathered are processed by a computer. https://en.wikipedia.org/wiki/Flow_cytometry Flow cytometry is routinely used in basic research, clinical practice, and clinical trials. Uses for flow cytometry include: Cell counting Cell sorting Determining cell characteristics and function

Detecting microorganisms Biomarker detection Protein engineering detection Diagnosis of health disorders such as blood cancers

	Measurable parameters https://en.wikipedia.org/wiki/Flow_cytometry#Measurable_parameters Apoptosis (quantification, measurement of DNA degradation, mitochondrial membrane potential, permeability changes, caspase activity) Cell adherence (for instance, pathogen-host cell adherence) Cell pigments such as chlorophyll or phycoerythrin Cell surface antigens (Cluster of differentiation (CD) markers) Cell viability Circulating tumor cells: isolation and purification Characterising multidrug resistance (MDR) in cancer cells Chromosome analysis and sorting (library construction, chromosome paint) DNA copy number variation (by Flow-FISH or BACs-on-Beads technology) Enzymatic activity Glutathione
Intracellular antigens (various cytokines, secondary mediators, etc.) Membrane fluidity Monitoring electropermeabilization of cells Nuclear antigens Oxidative burst pH, intracellular ionized calcium, magnesium, membrane potential Protein expression and localization Protein modifications, phospho-proteins Scattering of light can be used to measure volume (by forward scatter) and morphological complexity (by side scatter) of cells or other particles, even those that are non-fluorescent. These are conventionally abbreviated as FSC and SSC respectively. Total DNA content (cell cycle analysis, cell kinetics, proliferation, ploidy, aneuploidy, endoreduplication, etc.) Total RNA content Transgenic products in vivo, particularly the green fluorescent protein or related fluorescent proteins Various combinations (DNA/surface antigens, etc.)
Applications The technology has applications in a number of fields, including molecular biology, pathology, immunology, virology,[41] plant biology and marine biology.[42] It has broad application in medicine especially in transplantation, hematology, tumor immunology and chemotherapy, prenatal diagnosis, genetics and sperm sorting for sex preselection. Flow cytometry is widely applied to detect sperm cells abnormality associated with DNA fragmentation[43] in male fertility assays.[44] Also, it is extensively used in research for the detection of DNA damage,[45][46] caspase cleavage and apoptosis.[47] In neuroscience, co-expression of cell surface and intracellular antigens can also be analyzed.[48] In marine biology, the autofluorescent properties of photosynthetic plankton can be exploited by flow cytometry in order to characterise abundance and community structure. In microbiology, it can be used to screen and sort transposon mutant libraries constructed with a GFP-encoding transposon (TnMHA).[49]In protein engineering, flow cytometry is used in conjunction with yeast display and bacterial display to identify cell surface-displayed protein variants with desired properties. The main advantages of flow cytometry over histology and IHC is the possibility to precisely measure the quantities of antigens and the possibility to stain each cell with multiple antibodies-fluorophores, in current laboratories around 10 antibodies can be bound to each cell. This is much less than mass cytometer where up to 40 can be currently measured, but at a higher price and a slower pace. https://en.wikipedia.org/wiki/Flow_cytometry#Applications

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Antibodies

Immunostep has long been working to find and develop the best antibodies for Flow Cytometry. This has allowed us to obtain monoclonal antibodies that afford a better fluorescence intensity than other products available on the market. Moreover, Immunostep subjects all its antibodies to stringent testing such as repeatability, diagnostic sensitivity, analytical sensitivity or linearity, specificity and titre, thereby guaranteeing that all its products are suitable for diagnosis and research.

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