PASAP Hydrogel is a synthetic matrix used to create defined three-dimensional (3D) micro-environments for a variety of cell culture experiments. PASAP nanostructured hydrophilic hydrogel displays multiple functionalizations that fostering attachment, differentiation and viability of a variety of cells such as Neural stem cells, Mesenchymal stem cells, endothelial cells and fibroblasts.
PASAP provides the typical nanostructured topography of the extracellular matrix, thus mimicking the physiological microenvironment surrounding cells found in living tissues. This nanostructure supports cell attachment and spreading, thus fostering branching and differentiation: a paramount result of great importance to future therapeutic applications.
PASAP Hydrogel consists of a well-defined mixture of standard synthetic amino acids (1% w/v) and 99% water. Under physiological conditions, the peptide component self-assembles into a 3D hydrogel that exhibits a nano-fibrous structure (figure 1).
The hydrogel can be readily formed in culture dishes, multi-well plates, or cell culture inserts. PASAP applications include 2D and 3D cell cultures and in vivo cellular or drug therapies.
PASAP Hydrogel has been shown to promote survival (figure 2) and differentiation (figure 3) of murine and human Neural Stem Cells, of human Colon Stem Cells, of murine endothelial cells and of human fibroblasts. Preliminary studies also show that PASAP Hydrogel supports the in vivo engraftment of transplanted Neural Stem Cells into the site of acute Spinal Cord Injuries in Rats. Other potential applications include generic stem cell differentiation, tumor cell migration and invasion, angiogenesis assays, and in vivo therapies of tissue regeneration.
Figure 2. Murine Neural Stem Cells (A) and Human Neural Stem Cells (B) on PASAP Hydrogel in 2D cell culture conditions (7 DIV, 5x magnification). Murine Neural Stem Cells (C) and Human Neural Stem Cells (D) cultured in PASAP Hydrogel in 3D cell culture conditions (14 DIV): images are obtained by z-stack sectioning and flattening into single images (20x magnification in C, 5x magnification in D). Live Cells are marked in green. PASAP favors cell adhesion thus triggering stem cell differentiation and network formation. NSC cultured on untreated plastic wells do not attach and stay in round-shaped clusters.
Figure 3. Differentiated murine (A-B) and human (C-D) Neural Stem Cells in PASAP Peptide Hydrogel (96 MW) in 3D cell culture conditions (14 DIV): images are obtained by z-stack sectioning and flattening into single images. Cultures are serum-free and no animal-component has been used for tailoring the scaffold. Astrocytes and neurons are marked respectively in red (GFAP) and in green (bTubulin). PASAP fosters cell adhesion thus stem cell differentiation in networks of differentiating cell progenies. 10x and 20x magnification are respectively for A-C and B-D.
1% solution (w/v) of purified synthetic peptides
Packaged material exhibits pH = 3-5
Cell viability ≥ 80% based on cytotoxicity analysis of Neural Stem Cells.
Purity ≥99% confirmed using HPLC, LC-MS and MALDI-TOF.
Demonstration of fiber formation using self-assembly and rheology assays.
Purified Synthetic Peptide Composition
3D Hydrogel Structure