Edible Scaffolds Based on Non-Mammalian Biopolymers for Myoblast Growth

Abstract

In vitro meat has recently emerged as a new concept in food biotechnology. Methods to produce in vitro meat generally involve the growth of muscle cells that are cultured on scaffolds using bioreactors. Suitable scaffold design and manufacture are critical to downstream culture and meat production. Most current scaffolds are based on mammalian-derived biomaterials, the use of which is counter to the desire to obviate mammal slaughter in artificial meat production. Consequently, most of the knowledge is related to the design and control of scaffold properties based on these mammalian-sourced materials. To address this, four different scaffold materials were formulated using non-mammalian sources, namely, salmon gelatin, alginate, and additives including gelling agents and plasticizers. The scaffolds were produced using a freeze-drying process, and the physical, mechanical, and biological properties of the scaffolds were evaluated. The most promising scaffolds were produced from salmon gelatin, alginate, agarose, and glycerol, which exhibited relatively large pore sizes (~200 μm diameter) and biocompatibility, permitting myoblast cell adhesion (~40%) and growth (~24 h duplication time). The biodegradation profiles of the scaffolds were followed, and were observed to be less than 25% after 4 weeks. The scaffolds enabled suitable myogenic response, with high cell proliferation, viability, and adequate cell distribution throughout. This system composed of non-mammalian edible scaffold material and muscle-cells is promising for the production of in vitro meat.

Fabrication of Tissue Engineering Scaffolds Using Marine Bioactive Materials for Diverse Applications

“…The cells of other marine organisms can also be co-cultured with the scaffold for meat production. Enrione et al (2017) reported the growth of the Mus musculus muscle cells (C2C12) in the scaffold for meat production. A cost effective medium for the culturing of cells and identification of a plasticizer suitable for the type of cell used are necessary for the successful production of in vitro meat.…”

A New Edible Film to Produce In Vitro Meat

“…Prior to cell seeding, the films were soaked in CaCl 2 solution (70 mM) for 1 h to crosslink the alginate fraction and obtain a non-water-soluble material [8,24].…”

“…However, more basic research regarding optimization and production of muscle tissue for food products is necessary [7]. To make in vitro meat an economically viable food, it is necessary to investigate three key components, which are animal-free media, scaffolds [8], and bioreactors [9]. That is, methods to produce in vitro meat commonly employ the growth of myoblasts on a scaffold suspended in a serum-free culture medium in a bioreactor [3].…”

“…That is, methods to produce in vitro meat commonly employ the growth of myoblasts on a scaffold suspended in a serum-free culture medium in a bioreactor [3]. A scaffold is a matrix (soft material) where the anchorage-dependent cells (e.g., muscle cells) can adhere, remain viable, proliferate, and differentiate [4,8]. In addition, the use of mammalian components in the scaffold should be avoided to effectively reduce the slaughter of bovines [2].…”

“…In a prior work, we developed an optimized formulation to make edible scaffolds suitable for myoblast culture based on non-mammalian components [8]. The scaffold was formulated with a mix of three marine biopolymers (salmon gelatin, alginate, and agarose), where salmon gelatin performs as functional macromolecule containing RGD-sequences that promote cell adhesion and proliferation [12][13][14], alginate as crosslinker in contact with calcium, and agarose as gelling agent [8]. This edible scaffold was fabricated by freeze-drying, obtaining a microporous material where the myoblasts grew adequately, but the differentiation from myoblasts (muscle progenitor cell) to myocytes and finally muscle fiber were not researched.…”