Recipe for Recombinant Serum-Free Medium for Culturing Bovine Progenitor Cells
To create the serum-free medium for culturing bovine progenitor cells as described in Patent US20230083026A1 by Mosa Meat, you’ll follow a recipe that combines various components essential for cell growth and proliferation without the use of animal-derived products. This medium is particularly designed for bovine progenitor cells, including muscle and adipose tissue-derived cells, facilitating their expansion in an animal-free environment. Here’s a simplified recipe to prepare approximately 1 liter of the serum-free medium:
Ingredients:
- Recombinant Human Albumin: 5 g
- Fibroblast Growth Factor (FGF): 10 μg
- Ascorbic Acid (or derivative, e.g., L-ascorbic acid 2-phosphate): 50 mg
- Insulin: 10 mg
- Somatotropin: 2 μg
- Hydrocortisone: 36 μg
- Sodium Selenite: 6.7 μg
- Transferrin: 5.5 mg
- a-Linolenic Acid: 1 mg
- Ethanolamine: 2 mg
- L-Alanyl-L-Glutamine or Glutamine: 2 mM (or equivalent based on powder)
- Platelet-Derived Growth Factor (PDGF): 10 μg
- Insulin-like Growth Factor (IGF): 100 μg (optional based on specific needs)
- Vascular Endothelial Growth Factor (VEGF): 10 μg
- Hepatocyte Growth Factor (HGF): 5 μg
- Interleukin 6 (IL-6): 5 μg
- Basal Medium (DMEM/F12 or equivalent): Up to 1 liter
Equipment:
- Laminar flow hood for sterile handling
- Autoclaved glassware and pipettes
- Magnetic stirrer and stir bar
- pH meter
- 0.22 μm filter for sterilization
- Refrigerated storage
Preparation Steps:
- Sanitize Workspace: Clean and sanitize your workspace and all equipment using appropriate sterile techniques to prevent contamination.
- Prepare Basal Medium: Start with around 900 mL of your chosen basal medium (DMEM/F12) in a sterile container.
- Add Supplements: Sequentially add the powdered forms of recombinant human albumin, ascorbic acid, insulin, somatotropin, hydrocortisone, sodium selenite, transferrin, a-linolenic acid, ethanolamine, and glutamine into the basal medium. Use sterile techniques to transfer each component.
- Growth Factors and Cytokines: Add the fibroblast growth factor, PDGF, IGF (if used), VEGF, HGF, and IL-6. These are typically provided in liquid form, so pipette the necessary volumes into the medium.
- Adjust Volume: Add more basal medium to bring the total volume up to 1 liter.
- pH Adjustment: Check the pH of the medium. Adjust to the desired pH (typically around 7.2-7.4) using sterile HCl or NaOH.
- Sterilize: Filter the medium using a 0.22 μm filter to sterilize. Transfer the sterilized medium into a sterile container.
- Store: Label the medium and store it refrigerated (2-8°C) until needed.
Recipe for Recombinant Serum-Free Medium for Culturing Bovine Progenitor Cells
Usage:
- Gradually introduce your bovine progenitor cells to this medium, starting with a mix of their previous medium and this new formula, gradually increasing the proportion of the serum-free medium.
- Monitor cell adaptation closely, adjusting components if necessary based on cell response.
Note: This is a simplified guide based on the components listed in the patent. Actual preparation may require optimization based on specific cell line requirements and experimental conditions. Always consult material safety data sheets (MSDS) and wear appropriate personal protective equipment (PPE).
This recipe is derived from and inspired by the contents of patent US20230083026A1, titled “Serum-Free Medium for Culturing a Bovine Progenitor Cell,” filed by Mosa Meat B.V. The detailed descriptions and specific formulations provided in this recipe are based on interpretations of the patent document, which is publicly available for review. For commercial applications or further detailed study, please refer to the original patent document and consider consulting with a patent attorney or the patent holder for necessary permissions and licensing arrangements.
Tags: