Nanofibrous photoclickable hydrogel microarrays
Nanofibrous hydrogel microarray systems that act as facile, high throughput platforms for in vitro drug discovery and investigation and screening of combinatorial effects of physical and biochemical cues on maturation and differentiation of mammalian cells.
1. A method of high throughput combinatorial screening of engineered microenvironments comprising:
a. forming an in vitro cell culture matrix comprising:
i. a hydrogel fiber anchored to a solid support;
ii. biomaterial microdots arrayed on the hydrogel fiber; and,
iii. at least one mammalian cell seeded within the hydrogel fiber,
b. staining by immunofluorescence a protein in the at least one mammalian cell and
c. quantifying observed immunofluorescence from the immunofluorescence staining of the protein by microscopy and computer aided image acquisition, wherein the quantified immunofluorescence is a measure of at least one activity of the at least one mammalian cell selected from the group consisting of gene expression, cell function, metabolic activity, and cellular morphology.
2. The method of claim 1 , wherein the forming of the in vitro cell culture matrix comprises forming the hydrogel fiber by a fabrication technique selected from electrospinning, electrospraying, spin-coating, and deposition by dipping.
3. The method of claim 1 , wherein the biomaterial is at least one compound selected from a polysaccharide, a proteoglycan, a glycosaminoglycan, a cell membrane bound protein, a peptide signaling motif, a hormone, collagen I, collagen type II, collagen III, collagen IV, fibronectin, laminin, chitosan, elastin, entactin, fibronectin, tenascin, heparin sulfate, chondroitan sulfate, dermaten sulfate, and karatan sulfate.
4. The method of claim 1 , wherein the at least one mammalian cell is a stem cell.
5. The method of claim 1 , further comprising contacting the at least one mammalian cell with a test agent prior to measuring the at least one activity of the at least one mammalian cell.
6. The method of claim 5 , wherein the test agent is at least one of a growth factor, a hormone, a putative anti-cancer compound, a cell-surface protein inhibitor, a putative angiogenesis inhibitor, or a modulator of Epithelial-Mesenchymal Transition (EMT).