Beefy-9 Media — Experimentation to Discover a Cost-Effective Media for Bovine Stem Cells
Many cultivated meets have a successful proof of concept. We have cultivated meats like chicken, beef, and pork while other meats and animal-products are still in the process of being created. The real issue is how do we go from successfully creating these meats in labs to producing them in mass while also being cost-effective.
The Culprit: Cell-Media + Serum
Within Cellular Agriculture, researchers must use a media and serum to allow the bovine satellite cells (beef stem cells) to grow. Media is filled with salts, vitamins, carbon-based energy sources, and amino acids; aka the basics needed for a cell to grow. One such commonly used media includes Thermofisher’s Essential 8.
Half a liter of this medium runs for $245.00, leading to a net price per liter of $490.00 .
If $490/Liter seems like, just wait until you learn about FBS.
Serum; The actual culprit
In order to conduct successful bovine cultivation, researchers used a serum in addition to the media. A serum is comprised of electrolytes, antibodies, antigens, hormones, and lipids; aka all the stuff in blood, except the parts which may clot. The specific serum used is Fetal Bovine Serum (FBS). FBS is extracted from unborn fetal cows whose mother is recently deceased. Obviously, it’s not quite ethical, especially in the scope of Cellular Agriculture. What’s more shocking that the immorality of FBS is the price.
Media and serum usually amount for 55–95% of the net price fo create a cultivated meat. FBS alone can cost for more than $1000 a liter, in addition to the volatility of the serum.
Take for example, 500ml of FBS from Thermofisher costs around $803.00 . This would lead to a price per liter of around $1606.00 .
B8 Media
NewHarvest, a cellular agriculture research company, recently published a paper on a new, cheap, and effective media called B8. This new media does not need a serum (like FBS) and costs just around $16/L when producing the media in a lab.
B8 first originated to expand and grow Human induced pluripotent stem cells. B8, aka “Beefy-9”is comprised of Essential 8 medium, in addition to a basal media composed of glucose, amino acids, salts, amino acids, and vitamins, along with a few minerals and proteins.
NewHarvest grew bovine satellite cells (BSC), or cow stem cells, with varying rations of B8 and BSC-GM (typical media and serum) to see how media performs.
For the first experiment, NewHarvest tested B8’s efficacy in short-term growth of BSCs (3–4) days.
The x-axis displays the ratio of B8 media with BSC-GM, while the y-axis displays the level of fluorescence, a technique used to measure the amount of cells. Independently, neither BSC-GM or B8 show much growth in cells, but in a combination there’s a massive increase in cell population. Mixtures of B8 and BSC-GM showed a large increase in cell growth, thus proving the efficacy of B8.
Through this experimentation, we learned that B8 can reduce the need for BSC-GM (FBS Serum media), but it still has some faults. B8 media alone did not increase proliferation in proportion with the amount of B8, thus leading us to learn that the solution could be enhanced.
Media Supplementation
In order to see how this B8 + BSC-GM mixture could be improved, NewHarvest researchers tested out various acids and supplements to see how this new mixture could further increase growth of BSCs.
Supplements Tried:
- Interleukin (IL-6)
- Curcumin
- Recombinant Albumin
- Linoleic Acid
- Oleic Acid
Interleukin (IL-6)
Interleukin 6 (IL-6) is a type of interleukin, or a group of cytokines expressed by white blood cells. Interleukins have the job of mediating fever and fever reactions, especially the excess and lack of plasma reactions when fever is present. More appropriately, Interleukins have the ability to promote cell proliferation and differentiation.
As shown, IL-6 has the strongest increase in growth with 0.01 ng/mL. This shows how IL-6 is quite ineffective for BSC growth.
Curcumin:
Curcumin is produced from the Curcuma longa plants and takes shape as a yellowish powder for most of us. In a cellular perspective, curcumin is notable for inhibiting cell proliferation and cell apoptosis. In easy words, this means that the molecule doesn’t help much with cell growth as it inhibits cell death. This seems counterproductive towards the goals of cellular agriculture, something demonstrated by the experiment.
As predicted, Curcumin has little effect in improving cell growth, with increasing proportions of the molecule leading to a decrease in growth. 1 ng/ml revealed an increase in relative growth, but 100–5000 ng/ml demonstrated a loss in cell-growth.
Recombinant Albumin
- Recombinant — Relating to something who's DNA/Structure has been formed through recombination
- Albumin — Human protein created from the liver
Recombinant Albumin is a recombined protein, one naturally occuring from the human liver. Within our liver, albumin keeps the fluid within our bloodstream and prevents leaks. Albumin also carries materials such as enzymes, hormones, and vitamins — things also needed for BSC growth. In the scope of Cell-Ag, researchers would use Bovine Serum Albumin (BSA)(reminder, bovine = cow) to prevent adhesion of enzymes to tubes and pipettes, in addition to stabalize proteins. Recombinant Albumin, rAlbumin, should and does serve as a replacement for BSA.
Clearly, rAlbumin shows an increase in growth for BSCs. Rarely, this protein increases the growth almost in proportion to the amount of rAlbumin.
Linoleic Acid
Linoleic Acid is a fatty acid used for biosynthesis of prostaglandins and cell membranes. LA is a great for energy-storage and provides structure. LA is favorable for cell cultures as it’s insoluble in water and has been shown to enhance growth for other serum-free animal cell growth.
As for growing BSCs (Bovine Satellite Cells), Linoleic acid doesn’t quite reap as many benefits. Increasing amounts of LA lead to some growth, but 1600ng/mL and more harshly decrease the growth of BSCs.
Oleic Acid
Oleic Acid is an omega-9 fatty acid, something found commonly within food (all of these acids and supplements must be food safe because….we’re growing meat to eat it).
OA is quite similar to Linoleic acid, as it’s insoluble in water and also has shown to assist growth for other satellite cells.
When used as a supplement OA shows a very similar pattern to Linoleic Acid. There’s not much advantage, and then at 1600 ng/mL growth plummets to nearly 0.
Throughout these experiments, NewHarvest has shown how the new serum-free media, B8 combine with rAlbumin can significantly improve the growth and proliferation in the short-term timespan of 3–4 days.
While this does seem like a success, B8 has been demonstrated itself as a good starting point to work off of. Future experiments on the long-term success and efficacy of the media will prove whether or not this is a proper solution.
Thanks for reading! I’m Aryan Saha, a 16-year-old working to use technology to solve big problems. If you would like to contact me, reach out at aryannsaha@gmail.com. If you would like to connect, visit my linkedin or twitter
Sources:
https://www.biorxiv.org/content/10.1101/2021.05.28.446057v1.full#F1
