Engineering a 3D matrix from porcine plasma for enhanced cell proliferation and differentiation

The limited availability of suitable scaffolds for three-dimensional (3D) cell culture has driven the development of new biomaterials. In this study, we developed a porcine fibrin matrix gel (PFMG) derived entirely from porcine blood as a species-matched 3D scaffold that integrates the structural support of fibrin with the bioactive components of porcine platelet lysate (PPL).

PFMG was prepared from porcine plasma supplemented with PPL and calcium to induce gelation. Its physicochemical properties, including tunable stiffness, fibrillar microstructure, and molecular sieve-like behavior, were characterized by coagulation testing, diffusion assays, scanning electron microscopy, and rheological analysis. The biological performance of PFMG was evaluated using porcine mesenchymal stem cells (MSCs) and porcine embryonic fibroblasts (PEFs), and compared with conventional two-dimensional (2D) cultures supplemented with fetal bovine serum (FBS) or PPL. Cell proliferation, MSC tri-lineage differentiation, and transcriptomic changes were assessed.

PFMG formed a stable fibrin-based 3D network with adjustable mechanical properties and selective permeability to small molecules. Compared with conventional 2D culture systems, PFMG significantly enhanced the proliferation of both MSCs and PEFs and supported a characteristic 3D cellular morphology. MSCs cultured in PFMG retained adipogenic, osteogenic, and chondrogenic differentiation potential, and adipogenic differentiation was further enhanced when induction was performed with PPL instead of FBS. Transcriptomic analysis of MSCs cultured in PFMG revealed a distinct gene-expression profile characterized by upregulation of genes involved in cell-cycle progression, oxidative phosphorylation, and growth-related pathways, alongside downregulation of genes associated with cell adhesion and extracellular matrix interactions.

These findings indicate that PFMG provides a supportive, species-specific, and cost-effective 3D microenvironment for porcine cell culture. By combining scaffold function with platelet-derived bioactivity, PFMG promotes cell expansion while preserving multilineage differentiation capacity. This fully porcine platform therefore represents a promising biomaterial for porcine cell culture, tissue engineering, and regenerative medicine applications.