Our Technology
Role for hyaluronan in making primal mesenchymal stromal cells - Stromacytes
Our technology is based on our research on the role of hyaluronan in stem cell differentiation. This is a sugar polymer with wide spread roles in organ development, function and disease that depend on how it is presented to cells and other factors that interact with it. In our most recently peer reviewed study published in Stem Cell Research & Therapy, we report how it is sufficient to make distinctive primal mesenchymal stromal cells, we call Stromacytes, capable of modifying the function of blood cell lineages in the same way as adult organ derived cells.
Further investigations on Stromacytes
To understand how hyaluronan exerts its effect on stem cell differentiation we have deeply characterised cell gene expression following its presentation by our original and new methods versus the state of the art. This has informed distinctive changes induced by hyaluronan and co-factors, the nature of the primal cells which result, and the identity of molecules and pathways involved with potential to mediate regeneration of ageing cells and tissues.
Characterisation of cell gene expression by Principal Component Analysis of all expressed genes assessed by Total RNA sequencing in Stromacyte variants (SC 1.0, 2.0, 3.0) at designated passage (p) in culture, differing in how hyaluronan is presented by our original (1.0) and new intellectual proprietary (SC3.0 and +IP hPSC) vs state of the art (SC2.0 and -IP hPSC) methods applied to human pluripotent stem cells (hPSC), and also compared with human bone marrow mesenchymal stromal cells (BM MSC). Analysis performed using Qlucore Informatics Tools.
Stromacyte release proteins active in healing
In further investigations we have used Liquid Chromatography Mass Spectrometry (LCMS) and proteomic analysis to compare proteins released by variations of primal cells (Stromacytes) and adult bone marrow mesenchymal stromal cells into culture medium. This revealed that although there were differences in relative enrichment between cells, the same 400+ proteins appeared to be secreted by all cells. Bioinformatics analysis associated the activity of these proteins with molecular pathways involved in modulating immune cell function and extracellular matrix organisation involved in tissue healing.
Gene ontology network of Biological Process Pathways common to 400+ proteins represented by two or more unique peptides detected by LCMS in minimal chemically defined medium conditioned for 24 hrs with SC 1.o, 2.0, 3.0 and bone marrow mesenchymal stromal cells. Two pathways (nodes) are connected if they share 20% or more genes (proteins). Solid nodes are more statistically significantly enriched protein sets. Bigger nodes represent larger protein sets. Thicker edges represent more overlapped proteins. Gene Ontology networks calculated using ShinyGO 0.76
Released proteins enriched in Stromacytes
Variant primal stromacytes made by our hyaluronan based technology and other methods differ from each other and adult bone marrow mesenchymal stroma cells by relative enrichment of released proteins. For one variant these are distinctively enriched for activity in biological processes involved in cell, tissue and immune system formation and functions important in healing with ageing. So, primal cells have potential to mimic adult tissue mesenchymal stromal cells, and more functions supporting cell and tissue regeneration.
Proteomic analysis of proteins released by cells in culture. (A) Principal component Analysis of 400+ proteins secreted by Stromacyte variants (SC 1.0, 2.0, 3.0) vs bone marrow (BM) mesenchymal stromal cells showing distinctiveness of released protein profile for each type of cell as a whole. (B) Heat map of hierarchically clustered proteins showing enrichment for specific proteins (rows) in biological replicates of each condition (columns). Map is normalised to a mean equal to zero and variance equal to 1, Intensity of squares for each protein in each sample reflect log2fold increase (yellow) or decrease (blue) relative to mean (black). (C) Biological process pathways enriched in SC 3.0 released proteins. Two pathways (nodes) are connected if they share 20% or more proteins. Solid nodes are more significantly enriched protein sets. Bigger nodes represent larger proteins sets. Thicker edges represent more overlapped proteins. PCA and Heat Map prepared using Qlucore Informatics Tools. Gene Ontology networks calculated using ShinyGO 0.76