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
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.

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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.