Quantification of Cell Co-Migration Occurrences During Cell Aggregation on Fibroin Substrates

Abstract

A quantitative analytical method was proposed for measuring cell co-migration, which was defined as two or more cells migrating together. To accurately identify and quantify this behavior, cell migration on fibroin substrates was analyzed with respect to intercellular distance. Specifically, cell size was characterized by major diameter, and then, based on these measurements and cell center data, a specific threshold distance for defining co-migration was determined after analyzing cell motion using the Voronoi diagram method. The results confirmed that comigration occurrences of rounded cells were significantly more stable on fibroin than on ProNectin substrates under the present experimental conditions. The cell co-migration analysis method in this article was shown to be successful in evaluating the stability of cell co-migration and also suggested the presence of ''critical distance'' where two cells interact on fibroin substrates. With further research, the cell co-migration analysis method and ''critical distance'' may prove to be capable of identifying the aggregation behavior of other cells on different materials, making it a valuable tool that can be used in tissue engineering design.

Size dependent regeneration capacity of functionalized Capra ear-derived micro-tissue scaffolds for treatment of cartilage defects

“…In hanging culture model under gravitational force, MSs gradually settle down towards bottom forming cell-particle aggregates [17]. Inter-cellular distance has a major influence on aggregate formation since larger distance between cells inhibits inter-cellular cross talk [20]. Smaller particles ensure formation of compact clusters as the distance between particles is lower than the threshold distance for cell-cell communication, and cell density on smaller particles surface is also higher.…”

Silk fibroin sponges with cell growth‐promoting activity induced by genetically fused basic fibroblast growth factor

“…This is because, in addition to the high chondrocyte density observed on the bFGF‐fused fibroin sponges [Fig. (A,C)], the distinct adhesion and aggregation formation of chondrocytes observed on fibroin surfaces are considered to affect the chondrogenic phenotype positively . We are currently investigating the use of bFGF‐fused fibroin sponges as a scaffold for cartilage regeneration.…”

Higher Gene Expression Related to Wound Healing by Fibroblasts on Silk Fibroin Biomaterial than on Collagen

“…Wound dressings must protect and maintain suitably moist environments at the wound site to achieve effective healing throughout normal repair processes. Quantitative analyses of migrations and ECM productions of mouse fibroblasts on the SF surface revealed that fibroblasts on the SF surface migrated more actively and showed higher gene expression of ECM-related proteins than these on the collagen or fibronectin surfaces [12]. In the case of human keratinocytes, cells migrated more actively on the SF surfaces than on the Col surface.…”

“…Time-lapse microscopic images were taken using a high sensitivity cool CCD color camera (Keyence Co., Tokyo, Japan) for 24 h at 5 min intervals. Migration pathways of WI-38 fibroblasts on surfaces were tracked using MtrackJ software (http://www.imagescience.org/meijering/software/mtrackj/) for quantitative evaluation [12,13].…”

“…Our earlier work has demonstrated that chondrocytes cultured on SF sponges show enhanced synthesis of extracellular matrices (ECMs) and glycosaminoglycan, which are the main components of cartilage tissue [11,12]. The results show that more chondrocytes on SF adhered to the surface strongly, migrated, and aggregated than chondrocytes on glass or fibronectin surfaces [11,12].…”