Engineering multicomponent tissue by spontaneous adhesion of myogenic and adipogenic microtissues cultured with customized scaffolds

N Stephanie Kawecki; Sam C P Norris; Yixuan Xu; Yifan Wu; Ashton R Davis; Ester Fridman; Kathleen K Chen; Rachelle H Crosbie; Andrea J Garmyn; Song Li; Thomas G Mason; Amy C Rowat

doi:10.1016/j.foodres.2023.113080

Engineering multicomponent tissue by spontaneous adhesion of myogenic and adipogenic microtissues cultured with customized scaffolds

Food Res Int. 2023 Oct:172:113080. doi: 10.1016/j.foodres.2023.113080. Epub 2023 Jun 29.

Authors

N Stephanie Kawecki¹, Sam C P Norris², Yixuan Xu³, Yifan Wu⁴, Ashton R Davis⁵, Ester Fridman², Kathleen K Chen⁵, Rachelle H Crosbie⁶, Andrea J Garmyn⁷, Song Li⁸, Thomas G Mason⁹, Amy C Rowat¹⁰

Affiliations

¹ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Department of Materials Science and Engineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁴ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁵ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁶ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Neurology, David Geffen School of Medicine, University of California LA, USA; Broad Stem Cell Center, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁷ Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824, USA.
⁸ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA 90095, USA; Broad Stem Cell Center, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁹ Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Physics and Astronomy, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
¹⁰ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA 90095, USA; Broad Stem Cell Center, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: rowat@ucla.edu.

PMID: 37689860
DOI: 10.1016/j.foodres.2023.113080

Abstract

The integration of intramuscular fat-or marbling-into cultured meat will be critical for meat texture, mouthfeel, flavor, and thus consumer appeal. However, culturing muscle tissue with marbling is challenging since myocytes and adipocytes have different media and scaffold requirements for optimal growth and differentiation. Here, we present an approach to engineer multicomponent tissue using myogenic and adipogenic microtissues. The key innovation in our approach is the engineering of myogenic and adipogenic microtissues using scaffolds with customized physical properties; we use these microtissues as building blocks that spontaneously adhere to produce multicomponent tissue, or marbled cultured meat. Myocytes are grown and differentiated on gelatin nanofiber scaffolds with aligned topology that mimic the aligned structure of skeletal muscle and promotes the formation of myotubes in both primary rabbit skeletal muscle and murine C2C12 cells. Pre-adipocytes are cultured and differentiated on edible gelatin microbead scaffolds, which are customized to have a physiologically-relevant stiffness, and promote lipid accumulation in both primary rabbit and murine 3T3-L1 pre-adipocytes. After harvesting and stacking the individual myogenic and adipogenic microtissues, we find that the resultant multicomponent tissues adhere into intact structures within 6-12 h in culture. The resultant multicomponent 3D tissue constructs show behavior of a solid material with a Young's modulus of ∼ 2 ± 0.4 kPa and an ultimate tensile strength of ∼ 23 ± 7 kPa without the use of additional crosslinkers. Using this approach, we generate marbled cultured meat with ∼ mm to ∼ cm thickness, which has a protein content of ∼ 4 ± 2 g/100 g that is comparable to a conventionally produced Wagyu steak with a protein content of ∼ 9 ± 4 g/100 g. We show the translatability of this layer-by-layer assembly approach for microtissues across primary rabbit cells, murine cell lines, as well as for gelatin and plant-based scaffolds, which demonstrates a strategy to generate edible marbled meats derived from different species and scaffold materials.

Keywords: Cellular agriculture; Cultured meat; Scaffold; Tissue engineering; Tissue-engineered adipose tissue; Tissue-engineered skeletal muscle.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cell Differentiation
Gelatin*
Meat
Mice
Muscle Fibers, Skeletal*
Muscle, Skeletal
Rabbits

Substances

Gelatin

Grants and funding

F32 DE030004/DE/NIDCR NIH HHS/United States