La culture cellulaire

Résumé du document

Matériel requis

Pièce isolée
Hotte à flux laminaire
Incubateur
Refrigérateur
Congélateur
Aire de nettoyage
Matériel de culture
Eau distillée
Milieux de culture
Centrifugeuse
Microscopes
Haemocytomètre
Matériel de dissection
Système d'aspiration
Autoclave
(...)

Extraits

[...] Propagation: ATCC complete growth medium: Minimum essential medium (Eagle) with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate mM nonessential amino acids, and 1.0 mM sodium pyruvate, fetal bovine serum Temperature: 37.0 C Atmosphere: air, carbon dioxide (CO2) Subculturing: Protocol: 1. Remove and discard culture medium Briefly rinse the cell layer with Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. [...]

[...] Cells that are difficult to detach may be placed at 37°C to facilitate dispersal Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting Add appropriate aliquots of the cell suspension to new culture vessels Incubate cultures at 37°C Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended Medium Renewal: 2 to 3 times per week Freeze Medium: Freeze Medium: Complete growth medium supplemented with DMSO Storage temperature: liquid nitrogen vapor phase Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium) Bioreactive factors: Growth factor: T cell growth factor (TCGF); differentiation Le froid conserve Grâce à un cryoprotecteur les cellules peuvent être congelées et décongelées Le diméthylsulfoxyde DMSO 106 cellules/ml 10%DMSO 20%SVF Abaisser la température 1°C/min 1ère étape la glace 2ème étape 3ème étape Durée: de nombreuses années La décongélation doit être rapide min à 27 Les cultures spéciales Une lignée c'est bien mais 2 c'est encore mieux: La coculture Milieu de culture 28 En 2D c'est bien mais en 3D c'est encore mieux La matrice peut variée: collagène ou fibrine 29 2D contre 3D Le milieu de culture réalisées en 2D ou 3D polymérise avec du matrigel. [...]

[...] Four typical HeLa marker chromosomes have been reported in the literature. HeLa Marker Chromosomes: One copy of Ml, one copy of M2, four-five copies of M3, and two copies of M4 as revealed by G-banding patterns. M1 is a rearranged long arm and centromere of chromosome 1 and the long arm of chromosome 3. M2 is a combination of short arm of chromosome 3 and long arm of chromosome 5. [...]

[...] M3 is an isochromosome of the short arm of chromosome 5. M4 consists of the long arm of chromosome 11 and an arm of chromosome 19. Isoenzymes: G6PD, A Age: 31 years adult Gender: female Ethnicity: Black Hela Markers: Yes Comments: The cells are positive for keratin by immunoperoxidase staining. HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences. [...]

[...] Please click here for information regarding the specific requirements for shipment to your location. Related Cell Culture Products Applications: screening for Escherichia coli strains with invasive potential transfection host Virus Susceptibility: Poliovirus Adenovirus type Poliovirus Human poliovirus Encephalomyocarditis virus Reverse Transcript: negative DNA Profile Amelogenin: CSF1PO: 9,10 D13S317: D16S539: 9,10 D5S818: 11,12 D7S820: 8,12, THO1: 7 TPOX: 8,12 vWA: Cytogenetic Analysis: Modal number = 82; range = 70 to 164. There is a small telocentric chromosome in 98% of the cells aneuploidy in 1385 cells examined. [...]