DMEM/F-12 (with L-glutamine, sodium pyruvate, 15mMHEPES)

Product Description

DMEM/F-12 Medium (Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12) is a 1:1 mixture of DMEM and Ham’s F-12 medium. It is based on DMEM medium with the addition of nutrient-rich components from F-12 medium, including various trace elements. Dulbecco’s Modified Eagle Medium (DMEM) is a modified version of Eagle’s basal medium (BME), with four times the concentration of amino acids and vitamins compared to BME. Ham’s F-12, derived from Ham’s F-10 medium, significantly increases the concentrations of choline, inositol, putrescine, and several amino acids. DMEM/F-12 is widely used to support the growth of various mammalian cells, including MDCK, glial cells, fibroblasts, human endothelial cells, and rat fibroblasts. Additionally, DMEM/F12 serves as a foundation for developing serum-free culture media and is suitable for culturing mammalian cells under low serum conditions and at clone densities.

This DMEM/F-12 is modified as follows:

With	Without
L-Glutamine (365mg/L)
Sodium Pyruvate(55mg/L)
HEPES (3600mg/L)
Phenol Red(8.1mg/L)
Please refer to the instruction manual for detailed formulation.

Quality Control Index and Basis

• Appearance and Clarity (Appendix IXB of the Pharmacopoeia of the People’s Republic of China, 2010 Edition, Part II)
• pH value (Appendix VIH of the Pharmacopoeia of the People’s Republic of China, 2010 Edition, Part II)
• Osmolality (Appendix IXG of the Pharmacopoeia of the People’s Republic of China, 2010 Edition, Part II)
• Bacterial endotoxin (Appendix IXE to the Pharmacopoeia of the People’s Republic of China 2010 Edition Part II)
• Microbial Limit (Appendix IXJ to Part II of the Pharmacopoeia of the People’s Republic of China 2010 Edition)
• Cell growth test (Chemical Industry Standard of the People’s Republic of China “Mammalian Animal Cell Culture Media” HG/T3935-2007 Article 707)

Cell Growth Test

Test basis: Chemical industry standard of the People’s Republic of China “Mammalian animal cell culture medium” HG/T3935-2007 Article 707

Method:

1. Inoculate the cells into 6-well plates at 5×10^4/ml, 6 wells per cell per medium (3 wells for 48h assay + 3 wells for 48h change of media), the starting medium is the medium to be tested containing 5% FBS.
2. After 48h of static incubation in the incubator, cells from the 3 assay wells in each plate were digested and collected. And the cells were counted and viability was determined using a cell counter.
3. The medium in another 3 wells was discarded and FBS-free medium to be tested was added.
4. After continuing to incubate in the incubator for 48h, the cells in the fluid exchange wells were digested. Counting and viability assays were performed using a cell counter.

Stability Test

• Testing basis: industry standards and ICH guidelines
• Routine stability data: recommended shelf life of 12 months, respectively, in 2-8 ℃ storage conditions to test the stability of 3 months, 6 months, 12 months, 16 months, the results are not significantly different, in line with the quality control standards

Accelerated Destruction Test

The product was stored in an Electro-heating standing-temperature incubator and tested for stability at 37℃ and 45℃ respectively. At each temperature, divided into three groups: positive placement, inverted placement, horizontal placement; selected 8 hours, 24 hours, 48 hours, 72 hours node for testing. Product stability is good, no significant statistical differences.

Constant Temperature Transportation Simulation Test

The product was ordinary packaged by Shentong Express on July 12 from Dalian to Hainan, arrived on July 19th. Then immediately returned by Shentong Express, received on July 27th. Then tested, the product stability is good, no statistical difference, in line with the constant temperature transportation conditions.

Comparison of the results of cell culture medium with similar products form international brands

Cell	Brands	+5% FBS culture 48h		Change of medium, w/o FBS culture 48h
		Cell Number (pcs/ml)	Survival rate	Cell Number (pcs/ml)	Survival rate
Vero	G Brand	1.82 x10^5	95.3%	7.28 x10^5	93.5%
	MA0214	1.99 x10^5 ↑	95.7%	7.29 x10^5 ↑	93.4%
C6	G Brand	5.90 x10^5	94.1%	3.33 x10^6	95.7%
	MA0214	1.89 x10^5	94.5%	3.32 x10^6	95.6%
293T	G Brand	2.10 x10^5	95.6%	1.49 x10^6	96.9%
	MA0214	2.13 x10^5 ↑	95.7%	1.11 x10^6	94.9%
AGS	G Brand	1.47 x10^5	92.4%	2.38 x10^6	91.1%
	MA0214	1.73 x10^5	92.5%	2.38 x10^6	91.7%
L-292	G Brand	1.66 x10^5	92.0%	2.66 x10^5	87.4%
	MA0214	1.65 x10^5	93.8%	2.52 x10^5	87.0%
293	G Brand powder	1.03 x10^5	92.5%	1.72 x10^5	97.1%
	MA0214	1.03 x10^5 ↑	94.3%	2.11 x10^5 ↑	97.2%
SW480	G Brand powder	1.34 x10^5	89.6%	4.42 x10^5	92.5%
	MA0214	1.54 x10^5 ↑	91.4%	4.81 x10^5 ↑	92.7%
L-929	G Brand powder	1.41×10^5	92.5%	2.03×10^5	92.5%
	MA0214	1.53×10^5 ↑	92.8%	2.43×10^5 ↑	92.1%
Vero	G Brand powder	2.35×10^5	95.7%	1.15×10^6	98.3%
	MA0214	2.44×10^5 ↑	95.1%	1.14×10^6	98.5%

CONCLUSION: MeiunBio Basic Cell Culture Media is comparable to international first-class brands in terms of cell number and survival. Moreover, the cell morphology is good under microscope and the effect of long-term cell culture is stable, so it can perfectly replace the culture media of the first-line brand.

Shipping and Storage 

• Storage：Store at 2-8°C in the dark for one year.
• Shipment：Room temperature

Usage Statement

Research Use Only (RUO)