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| − | == Hela cells ==
| + | See wiki page for specific cell line |
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| − | == Muscle cells ==
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| − | In the Bioactuator project, we use C2C12 myoblasts. For more information about C2C12 myoblasts click on the following link: https://en.wikipedia.org/wiki/C2C12#:~:text=C2C12%20is%20an%20immortalized%20mouse%20myoblast%20cell%20line.&text=C2C12%20cells%20are%20used%20to,to%20explore%20mechanistic%20biochemical%20pathways.
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| − | We use the following cell culture protocol for the C2C12 cells:
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| − | '''C2C12 cells protocol'''
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| − | '''I-General information'''
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| − | 1. The base medium for this cell line Dulbecco's Modified Eagle's Medium (DMEM, Sigma D6429) with 10% fetal bovine serum (FBS) and 1% Penicillin/Streptomycin.
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| − | 2. The seeding density should be 5.0 X10<sup>3</sup>viable cells/cm<sup>2</sup>
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| − | 3. IMPORTANT - DO NOT ALLOW CULTURES TO BECOME CONFLUENT.
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| − | Cultures must not be allowed to become confluent as this will deplete the myoblastic population in the culture.
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| − | Myotube formation is enhanced when the medium is supplemented with 10% horse serum instead of fetal bovine serum.
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| − | '''II-Trypsinizing or passaging the cells'''
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| − | 1. Remove and discard culture medium. Rinse the cells with PBS twice.
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| − | 2. Add 2.0 mL of Trypsin-EDTA solution to flask and put it back in the incubator and wait about 5 minutes.Tap the flak and observe cells under an inverted microscope until cell layer is dispersed.
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| − | 3. Add 8.0 mL of complete growth medium and dispense cells by gently pipetting.
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| − | 4. Tranfer the cell suspension into 5a 0 ml tube.
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| − | 5. Centrifuge the cell suspension to make cell pellet (300g, 3 min). Remove the supernatant and re-suspend the cells in 10 ml of culture medium.
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| − | 6. Add appropriate aliquots of the cell suspension to new culture vessels.Inoculate at a cell concentration between 1.5 X 10<sup>5</sup> and 1.0 X 10<sup>6</sup> viable cells/75 cm<sup>2</sup> bottle.(Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
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| − | 7. Incubate cultures at 37°C.
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| − | 8. Medium Renewal: Every two to three days (see the medium colour)
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| − | == Collagen for 3D or coating surfaces ==
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| − | Type I Collagen is a major structural component of the extracellular matrix (ECM). Therefore, this fibrous protein is often used in three-dimensional (3D) collagen gels that simulate the in vivo cell environment better than the traditional 2D systems. Additionally, Collagen I is ideal for coating surfaces, as it can form thin layers for culturing cells.
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| − | [https://ibidi.com/cell-culture-microscopy/107-collagen-type-i-rat-tail.html#/110-concentration_volume-5_mg_ml_4_x_5_ml Type 1 Collagen from Ibidi], '''[https://ibidi.com/img/cms/products/cells_reagents/R_5020X_CollagenI/IN_5020X_CollagenI_05mg.pdf Protocols for use.]''' (bought through Inter Instrument https://interinst.no/. The product number is IB 50202 (4x5ml 5 mg/ml), price 3226,- + MVA Feb 2021).
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