Hyaluronic acid as a macromolecular crowding agent for production of cell-derived matrices

Sodium Hyaluronate Supplemented Culture Media as a New hMSC Chondrogenic Differentiation Media-Model for in vitro/ex vivo Screening of Potential Cartilage Repair Therapies

“…The early GAG production might also be explained by another interesting phenomena: macromolecular crowding. Macromolecular crowding (MMC), a biophysical phenomenon known to accelerate in vitro biological processes, accelerate and increases ECM deposition (Minton, 2001;Kumar et al, 2015;Shendi et al, 2019). The crowded environment is achieved by supplementation of the culture media with a synthetic or natural macromolecule (Kumar et al, 2015).…”

“…Thanks to the excluded volume effect exerted by the macromolecule, the culture media better recapitulates the crowded physiological in vivo environment and this is the reason of the in vitro acceleration of biological processes as ECM deposition (Kumar et al, 2015;Prewitz et al, 2015). Previous studies suggested that MMC can enhance GAG deposition during hMSC differentiation (Welter et al, 2009;Prewitz et al, 2015) and have shown culture media supplemented with 0.2% 1.8 MDa hyaluronic acid acts as macromolecular crowding agent (Zhou et al, 2008;Shendi et al, 2019). Therefore, the early sGAG production observed within the first week of chondrogenesis and particularly within the first 48 h of culture, could be explained by a potential macromolecular crowding effect exerted by hyaluronic acid supplemented in culture media.…”

Eclectic characterisation of chemically modified cell-derived matrices obtained by metabolic glycoengineering and re-assessment of commonly used methods

“…Thus, we consulted the literature and found that the majority of published studies on ECM determined the protein content using colorimetric assays such as the bicinchoninic acid (BCA) assay or the Bradford assay. 20,28,[42][43][44][45][46][47] These biochemical assays determine the total protein content in a solution through the interaction of the used dyes with the sample proteins leading to a photometrically measurable colour change of the sample solution. By comparing the measured absorbance of the protein sample to that of a standard protein of known concentration, the total protein content can be calculated.…”

Cardiac Cell-Derived Matrices Impart Age-Specific Functional Properties to Human Cardiomyocytes

“…The use of CDMs to study fundamental principles of cell-matrix interactions is an attractive approach that has increasingly grown in popularity in recent years 8,10,41,68 In addition to structural differences, CDMs had protein compositions specific to their developmental stage. Fetal CDMs were enriched for the three primary chains of Col 6, the loss of which is associated with impaired skeletal muscle function and regeneration due to metabolic deficits 62,63 .…”

“…Pre-treatment of tissue culture surfaces with the bioadhesive L-polydopamine covalently stabilized extracellular matrices following decellularization, while circumventing the need for adsorption of exogenous matrix proteins or complex surface chemistries and/or crosslinking. In addition, although MMCs have been used previously to enhance overall matrix production, characterization of MMC effects has been primarily limited to a few canonical ECM proteins such as Col I, FN and MMP241 . We show that the matrix produced in the presence of the MMC Ficoll is highly correlated with composition of matrix derived by CFs in the absence of MMCs.…”