Basics of Cell Culture

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Making a Primary <strong>Culture</strong>
Animal:PRIMARY CARDIOMYOCYTE CULTURE•newborn rats (Wistar)Heart isolation:•hearts were excised andcollected in sterile PBSDigestion:•after washing in fresh PBS, atria were removed and ventricules werecut into small pieces and put into the tripsin (0.25%) solution•digestion was proceed in 37 degree in the water bath for 25 minand the cell suspension was resuspended every 5 minutes using 5 mlpipettes
Page 1 and 2: Theoretical course: Basic biochemic
Page 3 and 4: Historical events in the developmen
Page 6 and 7: Continued...• 1965: Harris and Wa
Page 8 and 9: Why is cell culture used for?Areas
Page 10 and 11: Tissue culture• In vitro cultivat
Page 14 and 15: PRIMARY CARDIOMYOCYTE CULTUREWashin
Page 16 and 17: 2 days old neonatal cardiacmyocyte
Page 18 and 19: Adhesion• Majority Of Cells Adher
Page 20 and 21: Cell Surface Adhesion Molecules•
Page 22: Continous cell lines• Most cell l
Page 25 and 26: Culture media• Choice of media de
Page 27 and 28: Why sub culturing?• Once the avai
Page 29 and 30: Suspension cells• Easier to passa
Page 31 and 32: Freezing cells for storage• Remov
Page 33 and 34: Cell toxicity• Cytotoxicity cause
Page 35 and 36: Viability assay - LIVING CELLSCell-
Page 37 and 38: Hemacytometer• Specialized chambe
Page 39 and 40: Looking at the gridunder the phasec
Page 41 and 42: Viability assay - PI fluorescence i
Page 43 and 44: SIMULATEDISCHEMIA(Li et al 2004)CON
Page 45 and 46: Effects of Biological Contamination
Page 47 and 48: Basic equipments used in cell cultu
Page 49 and 50: Water bath• To warm media andPBS
Page 51 and 52: Mechanics of phase microscopyShifti
Page 53 and 54: Basic aseptic conditions• If work
Page 55: Reference:Paras Yadav 1 , Annu Yada

Basics of Cell Culture

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