Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway

HIC-5 expression was detected in osteosarkoma patients and osteosaroma cell lines by RT-PCR. Then RFP-SH-HIC-5 is transfected into the osteosarcoma cell line. The HIC-5 effect on cell viability, proliferation and apoptosis is assessed by MTT, Edu Kit and Flow Cytometry. Exosom is isolated from mg-63 cell supernatant by an exosomed insulation kit. Exosome-HIC-5 is confirmed by microscope electron transmission, particle size detection and RT-PCR.

Furthermore, the cells of the exosom-hic-5-5 are explored eligibility of cells, proliferation, and apoptosis. Furthermore, the C-IP test is used to identify the relationship between HIC-5 and SMAD4. TCF / LEF and the level of protein components of the WNT / β-Catenin signal are detected by the top luciferase test and Western blot. HIC-5 is regulated in osteosaroma tissue and cells. The decreased expression forced HIC-5 inhibits the proliferation of osteosarkoma cell lines, and induces the apoptosis of MG-63 and HOS. In Vivo, silencing HIC-5 sent the development of tumors. Furthermore, we isolate the exosom of the MG-63 supernatant, the excosom to conclude HIC-5 will regulate the proliferation and level of cell apoptosis of MG-63 and HOS. Furthermore, HIC-5 interacts with SMAD4 and regulates WNT / β-Catenin signals by reducing TCF / LEF activity.

Silencing HIC-5 inhibits proliferation and induces the apoptosis of cell osteosarcoma through deactivating the WNT / β-Catenin signal with the exosome path. Hypoxia is a characteristic of several serious diseases such as cardiovascular disorders and metabolism and cancer. A decrease at the oxygen level of the network induces hypoxic response in cells that seek to adapt to changing conditions. Failure to adapt to prolonged or severe hypoxia can trigger cell death. While some types of cells, such as neurons, are very susceptible to hypoxia, cancer cells utilize the hypoxia environment to undergo tumor growth, angiogenesis and metastasis. Hypoxic-induced processes trigger communication between complex cells and now there is an indication that the extracellular vesicle (EVS) plays a fundamental role in this process. The latest developments in EV isolation and the characterization methodology have raised awareness of the importance of EV purity in functional studies and cargo.

 

Exosomal limitations in the treatment of inflammatory diseases

Background: Despite great interest and many studies, there are currently no integrated standards that describe sequential manipulation with cells to get exosom for clinical use. The use of exosom has become an interesting alternative to cell therapy, because of the flexible properties of these biological vesicles. Enables scientists to manipulate their composition to produce the desired exosom to bring certain drugs, RNA and protein. This study aims to analyze the scientific literature on changes in the functional characteristics of exosom, depending on the manipulation method, which has the potential to contribute to the development of negative effects in the treatment of inflammatory genesis diseases. The choice of insulation method affects the set of protein markers disclosed, nucleic acid and receptors on microparticles.

Various surface receptors present on the exosome membrane can be engineered for the target of the lesion. Exocomals of healthy patients help reduce inflammation, normalize communication between cells and have antifibrotic, antioxidant and cytoprotective effects. Exosom can change the micro environment, but the micro environment can also change the composition of the exosomes.

Conclusion: Ecosomes obtained from sick patients carry markers of the characteristics of the appropriate disease. Such ecosomes can have pro-inflammatory properties, pro-fibrotic, cytotoxic, and oncogenic, and disrupt cellular cooperation. Until now, questions about doses, reactions to recurring giving, and the dose regime has not been fully resolved.

 Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway
Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway

Proteomic exploration based on mass spectrometry of small urinary extracellular vesicles in vasculitis associated with ANCA compared to total urine

The vasculitis associated with the ANCA (AAV) is a rare, but potentially serious self-immune disease, if not nowadays, with increased mortality and morbidity. Finding the first activity biomarkers and the prognosis is very important. The small extracellular vesicles (EVS) isolated from the urine can be considered as a non-invasive source of biomarkers. We evaluated several protocols for the isolation of urinary electric vehicles.

To eliminate contaminating non-vesicular proteins due to AAV associated proteinuria, we used the processing of proteinase K. We have studied the differences in proteomes of small patient events with AAV compared to healthy controls by the freedom of LC-MS / MS labels. In parallel, we performed a similar proteomic analysis of urine samples from identical patients. The results of the study showed significant differences and similarities in the EV proteome and urine, the latter being strongly affected by proteinuria. Using bioinformatic tools, we have explored the modified proteins differently and their related routes by focusing on AAV’s pathophysiology. Our results indicate a significant regulation of Golgi enzymes, such as MAN1A1, which may be involved in the activation of the T cell by N-glycans glycosylation and can play a key role in the pathogenesis and diagnosis of AAV.

Total RNA Isolation Kit II

FATRS--001 100 preps
EUR 369.6

Total RNA Isolation Kit II

FATRS--050 50 preps
EUR 259.2

Total RNA Isolation Kit (Plant)

P-9104 50 samples
EUR 416.7

Total RNA Isolation Kit (Tissue)

P-9102 50 samples
EUR 416.7

EpiQuik Total RNA Isolation Fast Kit

P-9105-050 50 samples
EUR 176

FastPure Cell/Tissue Total RNA Isolation Kit

RC101 50 rxn
EUR 274.8

FastPure Universal Plant Total RNA Isolation Kit

RC411-01 50 rxns
EUR 100

FastPure Cell/Tissue Total RNA Isolation Kit V2

RC112-01 50 rxns
EUR 82.6

Total RNA Isolation Kit II (Tri-reagent + RNA spin column)

FATRS__050 1 kit
EUR 62

Total RNA Isolation Kit II (Tri-reagent + RNA spin column)

FATRS__100 1 kit
EUR 112

FastPure Plant Total RNA Isolation Kit (Polysaccharides&Polyphenolics –rich)

RC401-01 50 rxns
EUR 100

FastPure Plant Total RNA Isolation Kit (Polysaccharides/Polyphenolics-Rich)

RC401 50 rxn
EUR 283.2

DNA/RNA Mini Kit (isolation of both genomic DNA and total RNA from a single sample)

FADRK_050 1 kit
EUR 90

Rna Isolation Kit

11828665001 EACH
EUR 378.62

Cartilage RNA Isolation Kit

K2031010 1 kit
EUR 443

RNA Broad Range Standard, 100 ng/uL

99850 1UL
EUR 63
Description: RNA from immortalized human cell line (Homo sapiens)

EVery EV RNA Isolation Kit

EVery100B-1 20 preps
EUR 412

Rapid Blood RNA Isolation Kit

BT4182 10Preps
EUR 98.36

Rapid Blood RNA Isolation Kit

BT4183 50Preps
EUR 197.03

Rapid Blood RNA Isolation Kit

BT4184 250preps
EUR 608.1

Rapid Plant RNA Isolation Kit

PT4191 50Preps
EUR 95.63

EpiMag Viral RNA Isolation Kit

P-9108-050 50 samples
EUR 152.9

Easy3-Plant RNA Isolation Kit

FYG502-100P 100 Preps Ask for price

Easy3-Plant RNA Isolation Kit

FYG502-500P 500 Preps Ask for price

Rapid Bacteria RNA Isolation Kit

BS8625 50Preps
EUR 95.63

AccuBlue Broad Range RNA Quantitation Kit (1000 assays)

31073 1kit
EUR 435.6
Description: Minimum order quantity: 1 unit of 1kit

AccuBlue Broad Range RNA Quantitation Kit (200 assays)

31073-T 1kit
EUR 169.2
Description: Minimum order quantity: 1 unit of 1kit

Reagent for Total Exosome Isolation (Culture Media Supplement)

EIR-02 50 ml
EUR 2485.2
Description: This product is used for isolating exosomes from culture media supplement based on chemical precipitation isolation method.

Phenol-Free Plant RNA Isolation Kit

10760056-1 50 Prep(s)
EUR 170.53

Easy3-RNA Isolation Kit

FYG501-100P 100 Preps Ask for price

Easy3-RNA Isolation Kit

FYG501-500P 500 Preps Ask for price

EpiQuik Magbeads Quick RNA Isolation Kit

P-9106-050 50 samples
EUR 163.9

Soil RNA Isolation Mini Kit (100prep)

FASRK-100 100 preps
EUR 224.4

Soil RNA Isolation Mini Kit (50prep)

FASRK-001 50 preps
EUR 180

EpiQuik Viral RNA Isolation Fast Kit

P-9107-050 50 samples
EUR 167.2

RNA-easy Isolation Reagent

R701-01 100 ml
EUR 201.6

RNA-easy Isolation Reagent

R701-02 200 ml
EUR 254.4

Easy3-Total NucleicAcid Isolation Kit

FYG504-100ML 100 ml Ask for price

Easy3-Total NucleicAcid Isolation Kit

FYG504-500ML 500 ml Ask for price

miRNA Isolation Kit (Tri-reagent + RNA spin column)

FAMIK__001 1 kit
EUR 134.5

miRNA Isolation Kit (Tri-reagent + RNA spin column)

FAMIK__002 1 kit
EUR 73

AccuBlue Broad Range dsDNA Quantitation Kit

31007 1KIT
EUR 374.4
Description: Minimum order quantity: 1 unit of 1KIT

PureXtract RNAsol RNA Isolation Solution, LS

R6103-120 2X60ml
EUR 343.2

Pharmalyte broad range pH 8-10.5 25mL

17045501 EACH
EUR 753.6

Pharmalyte broad range pH 3-10 25mL

17045601 EACH
EUR 630

Broad Range DNA Quantification Kit, 100 assays

NS1020-BR-100 1 each
EUR 131.3

miRNA Isolation Kit

FAMIK--001 Rxns: 100
EUR 398.4
Description: Contents of the kit:
  • Rxns:100Lysis
  • Buffer-10 ml
  • Acid-Phenol-10 ml
  • RNA Column and Collection Tube-100 sets
  • Wash Buffer-10 ml
  • Release Buffer-0.5 ml

miRNA Isolation Kit

FAMIK--002 Rxns: 50
EUR 267.6

miRNA Isolation Kit

FYG304-100P 100 Preps Ask for price

Pharmalyte broad range pH 2.5-5 25mL

17045101 EACH
EUR 753.6

Pharmalyte broad range pH 4-6.5 25mL

17045201 EACH
EUR 753.6

Pharmalyte broad range pH 5-8 25mL

17045301 EACH
EUR 753.6

Exosome Isolation Kit

E601E001 20 tests
EUR 2500

MicroRNA Isolation Kit

KS341025 1 kit
EUR 247

Endosome Isolation Kit

P528 - Ask for price

Microsome Isolation Kit

K249-50 each
EUR 633.6

Protoplast Isolation Kit

30210002-1 20 Prep(s)
EUR 199.16

Protoplast Isolation Kit

30210002-2 50 Prep(s)
EUR 438.32

CELLDATA RNAstorm™ Fresh Cell and Tissue RNA Isolation Kit, 50 preps

CD504 1
EUR 270
Description: DNAse I from bovine pancreas (cow; Bos taurus)

Chloroplast Isolation Kit

30210000-1 20 Prep(s)
EUR 273.97

Mitochondria Isolation Kit

AR0156 1 kit
EUR 127.2

Preadipocyte Isolation Kit

K583-5 each
EUR 639.6

Urine DNA Isolation Kit

K5011150 1 kit
EUR 237

Stool DNA Isolation Kit

K5011250 1 kit
EUR 237

Serum DNA Isolation Kit

K5018100 1 kit
EUR 355

AccuBlue Broad Range dsDNA Quantitation Solution

31009 1KIT
EUR 271.2
Description: Minimum order quantity: 1 unit of 1KIT

Prosi Prestained Protein Marker, Broad Range(11-170KDa)

P8500-040 200ulX2
EUR 271.2

RNA isolater Total RNA Extraction Reagent

R401-01 100 ml
EUR 159.6

Saliva DNA Isolation Kit

K5011050 1 kit
EUR 237

Genomic DNA Isolation Kit

55R-1362 50 assays
EUR 462
Description: Genomic DNA Isolation Kit for use in the research laboratory

Genomic DNA Isolation Kit

K281-50 each
EUR 326.4

Genomic DNA Isolation Kit

K2118-50 50 assays
EUR 385.2

AccuGreenâ„¢ Broad Range dsDNA Quantitation Kit (500 assays)

31069 1kit
EUR 267.6
Description: Minimum order quantity: 1 unit of 1kit

AccuGreenâ„¢ Broad Range dsDNA Quantitation Kit (100 assays)

31069-T 1kit
EUR 148.8
Description: Minimum order quantity: 1 unit of 1kit

Yeast Nuclei Isolation Kit

K289-50 each
EUR 470.4

Lipid Droplet Isolation Kit

MET-5011 50 preps
EUR 498
Description: Lipid droplets are lipid rich organelles found in the adipose tissue of eukaryotes. They function to regulate the hydrolysis and storage of neutral lipids and serve as storage for cholesterol and acyl-glycerols. Lipid droplets have also been associated with inflammatory responses, obesity, atherosclerosis, and cancer.

ExoPure? Isolation Kit   (Urine)

K1240-10 each
EUR 836.4

ExoPure? Isolation Kit   (Urine)

K1240-2 each
EUR 483.6

Apoptotic Cell Isolation Kit

55R-1349 30 units
EUR 874.8
Description: Apoptotic Cell Isolation Kit for use in the research laboratory

Apoptotic Cell Isolation Kit

K258-30 each
EUR 561.6

miRNA ISOLATION KIT-TISSUE

IB47370 4 PREP KIT
EUR 30.42

miRNA ISOLATION KIT-TISSUE

IB47371 100 PREP KIT
EUR 579.66

PSEUDOMONAS ISOLATION BROTH

P16-115-10kg 10 kg Ask for price

PSEUDOMONAS ISOLATION BROTH

P16-115-2Kg 2 Kg Ask for price

PSEUDOMONAS ISOLATION BROTH

P16-115-500g 500 g Ask for price

Exosome Isolation kit (for serum & plasma -Exosome-RNA-NGS)

P101NGS - Ask for price

Exo-Urine EV Isolation Kit

EXOU100A-1 2 x 5 reactions
EUR 490

FitAmp Gel DNA Isolation Kit

P-1007 100 Samples
EUR 324.48

Cell Mitochondria Isolation Kit

abx090630-50100tests 50-100 tests
EUR 309.6

Mitochondrial DNA Isolation Kit

55R-1361 50 assays
EUR 990
Description: Mitochondrial DNA Isolation Kit for use in the research laboratory

Mitochondrial DNA Isolation Kit

K280-50 each
EUR 705.6

Mitochondrial DNA Isolation Kit

K2117-50 50 assays
EUR 718.8

Cell Mitochondria Isolation Kit

ETF3601 100 assays
EUR 135

Mitochondrial DNA Isolation Kit

K2117-1 1 kit
EUR 858
Description: Isolates mt. DNA, no genomic DNA contamination

Whole Blood DNA Isolation Kit

K528-100 each
EUR 451.2

Plant/Fungi DNA Isolation Kit

10760114-1 50 Prep(s)
EUR 350.39

Yeast Mitochondria Isolation Kit

K259-50 each
EUR 502.8

FitAmp Urine DNA Isolation Kit 

P-1017
  • EUR 474.12
  • EUR 171.60
  • EUR 292.60
  • 100 Samples
  • 50 Samples
  • 100 Samples

XCF Exosomal DNA Isolation Kit

XCF200A-1 20 rxn
EUR 328

Tissue Mitochondria Isolation Kit

abx090631-50100assays 50-100 assays
EUR 309.6

Tissue Mitochondria Isolation Kit

ETF3606 100 assays
EUR 135

Lysosome Isolation Kit (20 tests)

P534 - Ask for price

Exosome Isolation kit (for urine)

P120L - Ask for price

Exosome Isolation kit (for urine)

P120S - Ask for price

FastPure FFPE DNA Isolation Kit

DC105-01 50 rxns
EUR 75.2

CD19 Positive Cell Isolation Kit

78564-1 1 x 10^8 Cells
EUR 70
Description: The CD19 Positive Cell Isolation Kit is designed to magnetically separate CD19-expressing-cells from a complex immune cell population. This kit is optimized for the isolation of CD19 positive cells from normal human peripheral blood mononuclear cells (PBMCs). Cells are incubated with the antibody:bead complex and placed on a magnet for quick and easy separation. When placed on the magnet, CD19-positive cells will be immobilized along the side of the tube while undesired CD19-negative cells will remain in suspension for easy removal.

CD19 Positive Cell Isolation Kit

78564-2 1 x 10^9 Cells
EUR 630
Description: The CD19 Positive Cell Isolation Kit is designed to magnetically separate CD19-expressing-cells from a complex immune cell population. This kit is optimized for the isolation of CD19 positive cells from normal human peripheral blood mononuclear cells (PBMCs). Cells are incubated with the antibody:bead complex and placed on a magnet for quick and easy separation. When placed on the magnet, CD19-positive cells will be immobilized along the side of the tube while undesired CD19-negative cells will remain in suspension for easy removal.

Genoirse Plant DNA Isolation Kit

GR101083 50
EUR 223

ExoPure? Isolation Kit   (Cell Media)

K1237-10 each
EUR 796.8

ExoPure? Isolation Kit   (Cell Media)

K1237-2 each
EUR 470.4

ExoPure? Isolation Kit   (Bio Fluids)

K1241-10 each
EUR 836.4

ExoPure? Isolation Kit   (Bio Fluids)

K1241-2 each
EUR 483.6

Lipopolysaccharide (LPS) Isolation Kit

K438-10 each
EUR 333.6

ExoPure? Isolation Kit   (Serum, Plasma)

K1238-10 each
EUR 836.4

ExoPure? Isolation Kit   (Serum, Plasma)

K1238-2 each
EUR 470.4

Apoptotic DNA Ladder Isolation Kit

K170-50 each
EUR 561.6

Cell Surface Protein Isolation Kit

K295-10 each
EUR 601.2

Apoptotic DNA Ladder Isolation Kit

K2045-50 50 assays
EUR 602.4

GPC1 Capture Exosome Isolation Kit

E601E004 20 tests
EUR 2500

CD63 Capture Exosome Isolation Kit

E601E005 20 tests
EUR 2500

CD81 Capture Exosome Isolation Kit

E601E006 20 tests
EUR 2500

Blood and Serum DNA Isolation Kit

K5017100 1 kit
EUR 355

Bacterial Genomic DNA Isolation Kit

K309-100 each
EUR 451.2

Breast cancer Exosome Isolation Kit

E601E002 20 tests
EUR 2500

EPCAM Capture Exosome Isolation Kit

E601E003 20 tests
EUR 2500

HSP70 Capture Exosome Isolation Kit

E601E007 20 tests
EUR 2500

EZExtract? Core Histone Isolation Kit

K234-100 each
EUR 314.4

10-min. Single Cell Isolation Kit

P713-20 - Ask for price

MagIso NK Cell Isolation Kit, Human

WHK-B023 20 tests, 200 tests Ask for price

Circulating Nuleic Acid Isolation Kit

FACFK-050 50 preps
EUR 589.2

Circulating Nuleic Acid Isolation Kit

FACFK-100 100 preps
EUR 990

Detergent-Free Nuclei Isolation Kit

P524-2 - Ask for price

Detergent-Free Nuclei Isolation Kit

P524-20 - Ask for price

Detergent-Free Nuclei Isolation Kit

P524-80 - Ask for price

Mouse Brain Microvessel Isolation Kit

P750-4 - Ask for price

Circulating Nuleic Acid Isolation Kit

FACFK_050 1 kit
EUR 476

MagIso CD3 Cell Isolation Kit, Mouse

WHK-B009 20 tests, 200 tests Ask for price

Org Frontier? Chloroplast Isolation Kit

K468-10 each
EUR 666

Meaning: The current study explores for the first time the changes in proteomes of small extracellular vesicles and urine of patients with vasculitis associated with former kidney relative to healthy controls by LC-MS / MS label. The isolation of vesicles from proteinuric urine samples has been modified to minimize plasma protein contamination and reduce co-insulation of extramuminal proteins. Differentially modified proteins and related routes with a role in AAV pathophysiology have been described and discussed. The results could be useful for the search for potential biomarkers in renal vasculitis associated with ANCa.