Influence of biological matrix and artificial electrospun scaffolds on proliferation, differentiation and trophic factor synthesis of rat embryonic stem cells

Abstract

Two-dimensional vs three-dimensional culture conditions, such as the presence of extracellular matrix components, could deeply influence the cell fate and properties. In this paper we investigated proliferation, differentiation, survival, apoptosis, growth and neurotrophic factor synthesis of rat embryonic stem cells (RESCs) cultured in 2D and 3D conditions generated using Cultrex® Basement Membrane Extract (BME) and in poly-(L-lactic acid) (PLLA) electrospun sub-micrometric fibres. It is demonstrated that, in the absence of other instructive stimuli, growth, differentiation and paracrine activity of RESCs are directly affected by the different microenvironment provided by the scaffold. In particular, RESCs grown on an electrospun PLLA scaffolds coated or not with BME have a higher proliferation rate, higher production of bioactive nerve growth factor (NGF) and vascular endothelial growth factor (VEGF) compared to standard 2D conditions, lasting for at least 2 weeks. Due to the high mechanical flexibility of PLLA electrospun scaffolds, the PLLA/stem cell culture system offers an interesting potential for implantable neural repair devices.

Bone marrow adipocytes in 3D

“…Pathway analysis of microarray data using the Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that genes involved in metabolic and proliferation pathways were upregulated in cells in 3D culture, whereas genes involved in inflammation and disease-related pathways were downregulated. These results agree well with previous observations that 3D cell culture favours the proliferation, differentiation and prolonged survival of cells 5,9 . It is well established that chronic low-grade inflammation, such as that present in obesity, leads to adipocyte and metabolic dysfunction.…”

Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins

“…Matrigel (BD Biosciences®) and Cultrex BME (Trevigen ®) are both biologically-derived matrices. They are reconstituted basement membrane extracts derived from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, and are rich in ECM constituents including laminin, collagen IV, perlecan, nodigen/entactin, proteases, as well as growth factors including epidermal growth factor (EGF) and insulin-like growth factor (IGF)[ 28 , 29 ]. It is noted that there is a difference in protein concentration between Matrigel and Cultrex BME, with typically 8–12 mg/mL in Matrigel [ 30 ] and 15–17 mg/mL in BME, respectively.…”

“…To our knowledge, there are no current studies that have compared the use of these matrices for 3D spheroids growth, despite there being a few studies that have noted the role of matrix in 3D cell growth. For example, it has been reported that rat embryonic stem cells (RESCs) grown in a synthetic poly-(L-lactic acid) (PLLA) scaffold proliferated at a faster rate compared to RESCs in BME [ 29 ]. Synthetic scaffolds support spheroid formation by providing a constructed porous structure for cells to grow into 3D structures inside the pores.…”

Biomimetic implants for pelvic floor repair

“…Hydrophilic materials have more affinity to water and adhesion proteins on the cell membrane . If a material is by nature hydrophobic, it can be blended with a more hydrophilic component, or one can use post‐modification methods such as plasma, laser, or grafting to increase the hydrophilicity. Hydrophilic or hydrophobic, electrospun matrices in general absorb more adhesion proteins than a non‐fibrous material because of their higher surface area‐to‐volume ratio and therefore, promote cell adhesion.…”