Decellularized Celery Scaffolds for Cell Culture and Guided Cell Alignment
New Harvest Research Fellow: Santiago Campuzano, BSc Food Science, University of British Columbia
Project Start Date: September 2017
Project Duration: Three years full time for a MSc
Institutes: University of British Columbia
Supervisors: Dr. Andrew Pelling (Canada Research Chair and Professor, Depts of Physics and Biology; University of Ottawa)
The goal of this work is to develop an open source, plant-based scaffolding platform which can be employed by anyone worldwide for numerous applications, including cellular agriculture.
Currently, biomaterial scaffolds that support the growth of mammalian cells in 3D are expensive and commonly derived from animal/human products, making them unsuitable for creating cultured meat.
The Pelling lab has demonstrated that apple-derived cellulose can act as an ultra-low cost and efficiently produced scaffold. It could support 3D culture of mammalian cells, promote cell invasion and proliferation, and retain shape and mechanical properties for several months in culture. As an organic, plant-based fibre, cellulose is one of the most abundant, sustainable, and easily sourced biomaterials on earth (it can be found in plant barks and leaves).
Over the following three years, Santiago will be investigating naturally-derived biomaterials from at least five plant sources — asian pear, carrot, rose petals, asparagus, and mushroom, to name a few — as scaffolds for cultured meat production.
Spatial alignment and orientation of cells in vivo, referred to as anisotropy, plays a crucial role in the functionality of tissue. The multinucleated structures in muscle tissue, known as myofibers, rely on uniaxial alignment to generate force along an axis. Airways, arteries, and veins rely on the circumferential alignment of smooth muscles to facilitate the transport of fluids and gases; and white matter in the brain relies on anisotropic axonal fibres for proper functioning. In the laboratory, however, 2D Petri dishes fail to recreate anisotropy. This in turn has shown a difference in gene expression, leading to unreliable results. To overcome this discrepancy, a wide array of methods, including topographical cues, cyclic strain, and electrical stimulation have been used to induce alignment. However, these methods are often laborious and rely on the use of specialized equipment. Consideration of recent publications on decellularized plant tissue as 3D substrates for cell culture has led us to speculate that a wide array of structures natively found in plants have yet to be explored. Here we depict the alignment of C2C12 murine myoblast on the decellularized vascular bundle of celery (Apium graveolens). The xylem channels (38.50μm ± 6.9) and phloem channels (21.5 μm ± 5.0) lie within the 10-100μm diameter necessary for optimal myoblast alignment. Following 10 days in proliferation media, the actin filaments of C2C12 and apex of nuclei were observed to be oriented parallel to the vascular bundle-grooves. Subsequently, after 5 days in differentiation media, myotubes with an average length of 308.1 ± 169.4μm (N=103) were -2.4° ± 3.8 (N=14) from the mean direction of the vascular bundle. We can therefore conclude that the microtopography of the vascular bundle guides muscle cell alignment. The results presented here highlight the potential of this plant-derived scaffold for in vitro studies of muscle myogenesis, where structural anisotropy is required to more closely resemble in vivo conditions.
Santiago at the microscope
Fun Facts: Santiago is into boxing and martial arts, worked in the hospitality industry as a cook and server throughout his degree, and just packed up his car for a 7-day cross-Canada move from Vancouver to Ottawa to begin his research.
This project is made possible thanks to the philanthropic support of the InVivo Group and the Scott and Cyan Banister Freedom Fund.