Fabrication of 3D microfluidic chip integrating concentration gradient design and perfusion system for embryo coculture with stromal cells

Fabrication of 3D chip that enables cell culture under continuous flow perfusion and repeated in situ observation by light microscopy is described. Thin polydimethylsiloxane (PDMS) membrane was etched to create an array of through-holes, and sandwiched between two PDMS channels to form an upper chamber and lower chamber. Thus we fabricated a 3D microfluidic chip which integrates the concentration gradient design and perfusion system for proposed embryo coculture with stromal cells.