MCDB 153 Medium is a low-protein and serum-free culture medium designed for specific cell types. MCDB 153 is one of the MCDB series media, designed and developed by Knedler and Ham. It was initially applied to the culture of human microvascular endothelial cells (HMVEC). In addition to endothelial cells, MCDB 153 medium is also suitable for other cell types, such as hepatocytes, smooth muscle cells, cardiomyocytes and so on.
Product Features:
MCDB 153 medium is developed for the serum-free culture of human keratinocytes. Before use, additives required for the growth of human keratinocytes need to be supplemented, such as recombinant epidermal growth factor (rEGF), bovine pituitary extract (BPE), hydrocortisone, etc.
Product Composition:
| Form | Liquid | | Concentration | 1× | | Specification | 500 mL | | pH | 7.2-7.4 | | L-Glutamine | 6mM | | NaHCO₃ | 1176 mg/L | | D-Glucose | 1100 mg/L | | Sodium Pyruvate | 55mg/L | | HEPES Buffer | Present | | Phenol Red Indicator | 12.4 mg/L |
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Operating Guidelines:
1. Warm trypsin (0.25% trypsin, 0.02% EDTA) and complete medium to room temperature in advance.
2. Discard the spent culture medium and rinse the cell monolayer with DPBS.
3. Add an appropriate volume of trypsin (for example, 5 mL for a 75 cm² culture vessel) to fully cover the cell monolayer. Incubate at 37 °C for 1-3 minutes. Observe under an inverted microscope until cells are completely detached from the surface of the culture flask. Gently tapping the flask can facilitate cell detachment.
4. Add 5-10 mL of complete medium, rinse the inner wall of the culture flask and mix well. Collect the cell suspension into a 15 mL centrifuge tube.5
5. Centrifuge at 1000 rpm for 5-10 minutes, discard the supernatant, and resuspend the cells with 2–5 mL of complete medium
6. Determine viable cell density and cell survival rate. Perform cell seeding, incubation and subculture following standard procedures according to cell type and experimental requirements.
Precautions:
1. Transport at room temperature. Upon receipt, store at 2- 8 °C in the dark and avoid repeated freeze-thaw cycles. The shelf life is 1 year.
2. This product is sterile filtered; gently invert and mix well before use.
3. Our company shall not be liable for any accidents caused by improper use of this product.
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