Micropatterned culture and differentiation of human bone marrow mesenchymal stem cells using a polydimethylsiloxane microstencil

A method for fabrication of polydimethylsiloxane (PDMS) microstencils was developed and its application to localized culture of human bone marrow mesenchymal stem cells (hMSCs) was tested. Unlike conventional culture methods, which culture cells on an entire surface, microscale cell culture provides precise control of the size and shape of stem cell patterns, and minimizes consumption of cells and culture media. A PDMS microstencil was fabricated by PDMS casting using an SU-8 mold prepared by photolithography. A pattern of 500-μm dots was tested. For the test, a PDMS microstencil was placed on a glass disk and cells were seeded on the stencil at a density of 5×10⁴ cells/cm². The hMSCs were cultured for 2 days at 37 °C in a humidified 5% CO₂ atmosphere. The PDMS microstencil was removed after 2 days and the hMSC patterns were inspected under a microscope. The results confirmed that stem cells can be cultured using a PDMS microstencil. The micropatterned hMSCs retained their ability to differentiate into osteogenic and adipogenic cells. Thus, using a PDMS microstencil, stem cells can be cultured and differentiated in micropatterns in a precisely controlled manner, in any shape and size, for research and bioengineering applications.