Using Microalgae Hydrolysates as an Alternative to DMEM and FBS in Cellular Agriculture

Team: Yomar Arnold, Elizabeth Walker, Britton Porter, Melanie Mills, Elise Barton

Sponsor: Upside Foods

Introduction

Cellular Agriculture is:

  • A new and quickly growing field in biological engineering
  • Attempting to reduce the need for and supplement current animal agricultural practices
  • Seeking an economical, serum-free cell growth media
  • Cellular Agriculture uses less energy, water, land, and produces fewer greenhouse gas emissions than traditional animal agricultural practices
  • Current culture media is too expensive for feasible scale up
Figure 1. Timeline of Cellular Agriculture in the United States

Figure 1. Timeline of Cellular Agriculture in the United States

Objectives

  • Reduce the cost of cell culture media by 20%
  • Utilize both common and uncommon feedstocks for media:
  • Chlorella vulgaris, Arthrospira platensisboiled extracts
  • Wheat, Peptone, Soy, and Yeast hydrolysates
  • Maintain cell growth and morphology
  • Optimize ratios of extracts and hydrolysates to result in the best cellular yields
  • Adhere to the Generally Recognized as Safe (GRAS) guidelines for hydrolysate feedstock selections

Background

Figure 2. The basic cellular agriculture process

Figure 2. The basic cellular agriculture process

Methods

Algal Extracts

Figure 3. Method of creating algal extracts

Figure 3. Method of creating algal extracts

Hydrolysates

All other hydrolysates used were either purchased premixed, or were mixed to a 10% solids slurry w/w and were sterilized by syringe filtration

Cell Culture

Figure 4. Cell culture and passaging technique

Figure 4. Cell culture and passaging technique

Results

Figure 6. Average cell counts normalized to cells grown in DMEM, 95% CI

Figure 6. Average cell counts normalized to cells grown in DMEM, 95% CI

Media Cost w/o FBS (USD) Cost w/ FBS (USD) Percent Cost Was Reduced w/ FBS
DMEM (10% FBS) 1,340,000 5,140,000 0.00
Peptone 23,600 811,000 84.22
Yeast 56,200 850,000 83.46
Spirulina 7,500 800,000 84.44
Chlorella 13,000 801,000 84.42
Spir/Wheat/Chlor 30,000 820,000 84.05
Soy/Wheat 3,900,000 4,600,000 10.51
Yeast/Chlor 35,000 822,000 84.01
Yeast/Spir 32,000 820,000 84.05
Pep/Chlor 18,000 806,000 84.32
Pep/Spir 16,000 803,000 84.38
Soy/Wheat/Spir 2,600,000 3,400,000 33.85
Chlor/Spir 10,000 798,000 84.47
Soy/Wheat/Chlor 2,630,000 3,360,000 34.63

Figure 7. Approximate cost of each media ratio, and reduction (FBS was at 2% v/v unless noted)

Conclusion

  • Microalgae hydrolysates show promise as an alternative to DMEM and FBS in Cellular Agriculture
  • Based on cost of media, microalgae also shows promise as a more economical alternative to DMEM and FBS
  • Further research is needed using a cell line meant for human consumption instead of HEK Cells