1. Technique Cell culture and maintenance of cell lines: 1.1.- Introduction and working rules. 1.2.- Culture of pre-adipocytes: preparation of complete DMEM culture media. Preparation of specific culture media for adipocyte differentiation and maintenance. 1.3.- Freezing (preparation of freezing medium, cryovials). Thawing of cells. 1.4.- Cell seeding, confluence, calculation of cell density - flasks and P96, P12 and P24 plates. Cell passages. 1.5.- Automated cell counting (trypan blue staining). 1.6.- Operation and use of general culture laboratory equipment. 1.7.- Biological safety (ethanolisation, sterility, washing of culture cabinets, glass and plastic residues).2. Extraction and quantification of RNA by spectrophotometry and assessment of its purity. 2.1.- Maintenance of the RNA extraction area. 2.2.- Use of NanoDrop. Absorbance.3. RT-PCR technique. 3.1.- Calculation of the volume of cDNA from the RNA obtained. 3.2.- Retrotranscription. 3.3.- Plates and plate design for PCR. 3.4.- Thermocycler.4. Quantification of intracellular fat. 4.1.- Triglyceride staining (Oil Red O). 4.2.- Spectrophotometry.5. Quantification of cell viability. 5.1.- MTT assay. 5.2.- Spectrophotometry.6. Adipocyte differentiation stage. Photomicroscopy. 7. Organisation and time optimisation: laboratory notebook.8. Excel.

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