Features of Microsystems for Cultivation and Characterization of Stem Cells with the Aim of Regenerative Therapy

Figure 1

(a) Fabrication of single HSCs microwell array. (i) Reactive thiol and vinyl sulfone end-groups on poly(ethylene glycol) (PEG) precursors. (ii) Overview of hydrogel microwell array fabrication. A PDMS stamp is cast on a silicon master (step 1). This stamp is used as a template to crosslink a PEG gel containing the complementary microwell array topography (steps 2 and 3). Upon swelling and washing, the hydrogel surface is used to trap individual HSCs (step 4). Typical dimensions of microwells are indicated. (b) (i) Coculture system using parylene-C stencils. For cell culture on PDMS, fibronectin was coated on the PDMS surface. (ii)~(v) Light micrograph (left) and the corresponding fluorescent (right) images of the steps in the formation of dynamic cocultures using parylene-C stencils. (ii) Hyaluronic acid-coated parylene-C stencil was reversibly sealed on fibronectin treated PDMS and seeded with mES cells. (iii) The patterned cocultures of mES cells and AML12 hepatocytes. (iv) To generate dynamic cocultures, the stencil was gently peeled away, leaving the mES cells. (v) After depositing a layer of fibronectin, a third cell type (NIH-3T3) was seeded on the exposed surface. Scale bars are 200 μm.

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