Simplified low-cost methodology to establish, histologically process and analyze three-dimensional cancer cell spheroid arrays
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Simplified low-cost methodology to establish, histologically process and analyze three-dimensional cancer cell spheroid arrays
Different from the two-dimensional cell culture, three-dimensional (3D) model of multicellular spheroid allows cells to build interactions cell-cell / cell-matrix on the surface of cells throughout, more closely mimic microenvironments of tumors and a subpopulation cell with a certain standard morphology, differentiation and expression gen.
Since then several methodologies that involve or aggregate 3D generation cells or histological processing and analysis have appeared, but in general they are laborious, expensive and complicated to set up as a routine technique. Thus, we developed a complete methodology, detailing simple, accessible and inexpensive step by step, including 1) 3D aggregate generation cells using the technique of reduction depends; 2) provide a simple way to assess the morphological parameters of spheroids produced; followed by 3) multiples and organized histological processing, keeping some individual spheroids in an agarose tool, maintaining order is known and the position of each, similar to a tissue microarray principle; 4) until the last step, in which he allowed the simultaneous histological analysis of the composition of a few slices of round, well-organized side by side, in the same block, through conventional staining or 5) immunostaining against different molecular markers.
Therefore, the purpose of present methodology for the popularization of 3D cell culture, enabling to make these unusual techniques in basic cell biology research, after all the steps performed without the use of heavy reagents, materials or equipment. Besides bringing the apparatus simple agarose as new low cost, which allows high-throughput analysis of multiple spheroids simultaneously in an organized manner.
Simplified low-cost methodology to establish, histologically process and analyze three-dimensional cancer cell spheroid arrays
Synthesis and Evaluation of Some Novel Antimicrobial pyrimidine, pyrazole, Chromene and Tetrahydrobenzo [b] thiophene derivatives Bearing Pyrimidinthione part
Aims and objectives: A collection of novel fused pyrimidine, pyridine, pyrazole, chromene, thiophene derivatives 2-30 has been newly synthesized by using 1a, b as the starting material. Fused Pyrans show off a variety of pharmacological activities such as cancer agents [1], antimicrobial [2-4], antioxidants [5], the anti-proliferation [6], cytotoxic activity [7], the anticipated anti-tumor [8], antiparkinson [9] and anti-inflammatory [10]. Also, derivatives Pyrans famous for bacterial biofilm disruptor [11], anticonvulsants [12] and inhibtors of mycobacterium bovis [13].
Materials and methods: All melting points are measured using Block Akofler instrument and are uncorrected. IR spectrum (KBr) recorded in 5300 FTIR spectrometer (υ, cm-1). 1H-NMR were recorded on a Varian Gemini spectrometer. 1H-NMR run at 300, 400 MHz and 13 C-NMR run at 100 MHz in DMSO-d6, CDCl3 as solvent.
Chemical shifts expressed in parts per million (ppm) using tetramethylsilane (TMS) as an internal refernce. 1000 EX mass spectrometer at 70 eV. The purity of the synthesized compounds is checked by thin layer chromatography (TLC) (aluminum sheet) using n-hexane, EtOAc (9: 1, V / V, 7: 3 V / V) eluent. Elemental analysis performed by the microanalytical Research Center, Faculty of Science, and microanalytical Unit, Faculty of Pharmacy, Cairo University, Egypt.
Description: A sandwich quantitative ELISA assay kit for detection of Human Carbohydrate Antigen 15-3 (CA15-3) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Human Carbohydrate Antigen 15-3 (CA15-3) ELISA Kit
Description: A sandwich quantitative ELISA assay kit for detection of Human Carbohydrate Antigen 15-3 (CA15-3) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Human Carbohydrate Antigen 15-3 (CA15-3) ELISA Kit
Description: IL-15 is an immunomodulating cytokine that stimulates the proliferation of T lymphocytes and shares many biological properties with IL-2. IL-15 exerts its biological activites primarily on T cells. It is also essential in the development, survival and activation of NK cells. Increased expression of IL-15 has been implicated with rheumatoid arthritis, inflammatory bowel disease and diseases affiliated with retroviruses HIV and HTLV-I. Human IL-15 is biologically active on mouse cells as measured by the dose-dependent stimulation of the proliferation of mouse CTLL cells. Recombinant human IL-15 is a 12.9 kDa protein consisting of 114 amino acid residues.
Description: Interleukin-15 (IL-15) is a cytokine with structural similarity to IL-2 that is secreted by mononuclear phagocytes following infection by virus(es). This cytokine induces cell proliferation of natural killer cells. Swine IL-15 Recombinant Protein is purified interleukin-15 produced in yeast.
Description: Cancer Antigen 15-3 MUC 1 Antigen, Host/Source: Human Milk. The purity is detected by salt extraction and delipidization following high speed centrifugation. It is tested negative for HBsAg, antibodies to HCV and HIV 1/2.
Description: Our tissue products are produced by strictly following the IRB ethical standards and procedures and from highest quality tissues. Immediately after collection the tissues are placed in liquid nitrogen and examined by certified pathologists. The thickness of each individual section is ~5um. They are Hematoxylin and Eosin stained and quality tested by immunostaining with anti-beta-actin antibodies. Our tissue products are suitable for various studies on cellular level (RNA localization, Protein expression, etc.) on both normal and pathological cases. It is also an excellent control and educational tool.
Description: Interleukin-3 Human Recombinant produced in E.Coli is single, a non-glycosylated, Polypeptide chain containing 154 amino acids fragment (20-152) and having a total molecular mass of 17.3kDa and fused with a 20 aa N-terminal His tag. ;The IL3 His is purified by proprietary chromatographic techniques.
Description: TGFB3 Human Recombinant produced in plant is a disulfide-linked homodimeric, glycosylated, polypeptide chain containing 118 amino acids and having a molecular mass of 27.2kDa. ;The TGFB3 is fused to 6xHis tag at N-terminus and purified by standard chromatographic techniques.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: Matrix metalloproteinases (MMPs) are a family of endoproteases that require zinc and calcium for expressing catalytic activity. These enzymes play a central role in the maintenance and remodeling of the extracellular matrix. Elevated expression of their activity, caused either by up-regulation of their expression or down-regulation of their cognate inhibitors, has been implicated in various degenerative disorders, including arthritis, cardiovascular disease, skeletal growth-plate disorders, and cancer metastasis. MMP-3 degrades fibronectin, laminin, collagens III, IV, and X, and cartilage proteoglycans. Recombinant human MMP-3 is a 42.8 kDa protein containing the entire catalytic N-terminal domain and the C-terminal domain (378 amino acids).
Results: A series of novel azoles and azines designed and prepared by reacting 7-amino-5- (4-chlorophenyl) – 4-phenyl-2-thioxo-2,5-dihydro – 1H pyrano- [2, 3-d] pyrimidine -6-carbonitrile 1a and 7-amino-4,5-diphenyl-2-thioxo-2,5- dihydro-1H-pyrano [2,3-d] -pyrimidine-6-carbonitrile 1b with some electrophilic and nucleophilic reagents. The structure of the target compounds were confirmed by elemental analysis and spectral data. Newly synthesized compounds showed good antimicrobial activity against microorganisms mentioned earlier.
