293 Serum-free Culture Medium (with L-Glutamine)

Item Number

S743060

• Stability And Storage: Store at 4℃ in the dark for up to one year.
• Storage Temp: Store at 2-8°C,Protected from light
• Shipped In: Wet ice

Product Description

Aladdin's 293 Serum-free Culture Medium (with L-Glutamine) is a ready-to-use, chemically defined liquid culture medium specifically developed and optimized for the high-density growth of various HEK293 cells including 293S, 293F, as well as domesticated 293H, 293E, 293T [1-3]. It is free of protein, growth factor, and substances of animal origin.The 293 Serum-free Culture Medium is also known as CD (Chemical-defined) 293 Medium. Human Embryonic Kidney (HEK) 293 cells are derived from human embryonic kidney cells [4]. This product containing L-Glutamine at a concentration of approximately 6mM has been sterilized by filtration and can be directly used for serum-free culture of 293 cells. Additional supplementation of L-Glutamine, serum, antibiotics, non-ionic surfactants (such as F-68), and anti-clumping agents is generally not required.Component

Application：

This product can support high-density growth and efficient transient expression in various 293 suspension cells, such as 293S and 293F. It is widely used in high-efficiency packaging or amplification of viruses, production of recombinant proteins, and large-scale antibody production based on cell culture. It can also be used for other domesticated 293 cells, subtypes, or derived cell lines, such as domesticated 293H, 293E, and 293T.

Precautions：

When storing this product at 4℃, do not place it near the walls of the refrigerator to avoid precipitation. If precipitation occurs, gently shake the product at room temperature for a moment, then place it in the refrigerator at 4℃ overnight or longer until the precipitation disappears before use.This product is developed for 293 suspension cells and is not recommended for non-domesticated 293 adherent cells.This product should be handled aseptically to avoid microbial contamination. It is generally not recommended to supplement with antibiotics as it may affect the normal growth of cells. However, if it is not possible to operate in a highly sterile environment, an appropriate amount of antibiotics may still need to be added.When turbidity or precipitation occurs in the culture medium during use, it should be discarded.The L-Glutamine (approximately 6mM) contained in this product is an essential nutrient for cell culture and does not usually require additional supplementation. However, as the L-Glutamine is not very stable and easily decomposes in solution, the culture medium that has been stored for a long time may cause slow cell growth. Under such a circumstance, an appropriate amount of L-Glutamine (100X) (, C0212) or L-Ala-Gln (100X) (, C0211) can be added before use to maintain normal cell growth and metabolism.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1.Recovery of 293 Suspension Cellsa.Take out the cell cryovial from liquid nitrogen or a ultra-low temperature freezer and rapidly thaw it completely in a 37℃ water bath (keep the mouth of the cryovial above the liquid to avoid contamination). Gently swirl the cryovial to facilitate thawing, but do not vortex. Rapid and complete thawing can improve cell recovery.b.Before opening the cryovial, wipe the external surface of the cryovial with 70% ethanol. Be careful not to wipe off the markings on the cryovial.c.Centrifuge the completely thawed cells directly or transfer them to a sterile 1.5ml or other suitable centrifuge tube. Centrifuge at 200×g for 5 minutes at room temperature and remove the supernatant carefully without disturbing the cell pellet.d.Resuspend the cells in the 293 Serum-free Culture Medium pre-warmed at 37℃ and transfer them to a culture vessel (such as a conical flask or a triangular cell culture flask) made of PC or PETG material. Mix well and place the culture vessel in a cell culture shaker at 37℃, 5-8% CO2, and 80% humidity. Set the shaker speed to 130rpm (with an amplitude of 26mm). Note: If the shaker has an amplitude of 50mm, set the speed to 120rpm.2.Passaging of 293 Suspension Cellsa.Passage 293 cells that are in the logarithmic growth phase, with a cell viability greater than 95% and a cell density of 2-4×106 cells/ml or higher.Note 1: The logarithmic growth phase may vary for different types of 293 cells.Note 2: Cells that have been passaged 30 times or for more than 3 months should not be used for more passages.Note 3: We recommend using Transfection Reagent or Linear Polyethylenimine specifically developed for transfection of 293 suspension cells for transfection according to manufacture’s instructions. b.Collect 293 suspension cells in logarithmic growth phase, with cell viability ＞95% and a density of 2-4×106 cells/ml for transfection.c.Dilute 0.7-2mg of plasmid DNA in 10ml of 293 Serum-free Culture Medium, mix well gentlyd.Dilute an appropriate amount of PEI in 10ml of 93 Serum-free Culture Medium, mix well gently.e.Add the diluted PEI to the diluted plasmid DNA, gently invert 4-5 times to mix thoroughly, and incubate at room temperature for 15-20 minutes.Note: The order of mixing the PEI with the plasmid DNA should not be reversed.f.Add the mixture to the cells dropwise while gently rocking the triangular cell shaker during the addition.g.Place the cells in a cell culture shaker under normal cell culture conditions. Typically, cells can be maintained in this 293 Serum-free Culture Medium for 48 hours without the need for additional supplements. Alternatively, after transfection, add a certain amount of supplements to the cells every 24 hours while gently rocking the culture container during the addition. Afterwards, return the cells to the cell culture shaker for continued culture.h.Collect the cell supernatant or cells for downstream applications when the cell viability is ＜60%.References：1.Schneider MD, French BA. Circulation. 1993. 88(4 Pt 1):1937-42.2.Fisher KJ, Jooss K, Alston J, Yang Y, Haecker SE, et al. Nat Med. 1997. 3(3):306-12.3.Yan SB, Chao YB, van Halbeek H. Glycobiology. 1993. 3(6):597-608.4.Graham FL, Smiley J, Russell WC, Nairn R. J Gen Virol. 1977. 36(1):59-74.Related Products:Cat. No.Product NamePack Size C0212 L-Glutamine (100X) 100ml C0211-100ml L-Ala-Gln (100X) 100ml C0901-500ml 293 Serum-free Culture Medium (with L-Glutamine) 500ml C0901-1L 293 Serum-free Culture Medium (with L-Glutamine) 1L C0537-20mg Polyethylenimine, Linear 20mg C0537-100mg Polyethylenimine, Linear 100mg C0518-1ml Lipo293F™ Transfection Reagent 1ml C0518-10ml Lipo293F™ Transfection Reagent 10ml C0518-100ml Lipo293F™ Transfection Reagent 100ml