Microstencils have been utilized in biomedical engineering to pattern cell cultures, engineer tissues, and pattern conductive materials in microelectronics for the measurement of bioelectric activity. However, fabricating these microstencils can be considerably time consuming, expensive, and cleanroom processing or laser micromachining intensive. We present microfabrication strategies for producing stencils rapidly and cost effectively, with minimal use of cleanroom facilities, ideal for prototyping or applications where microfabrication costs need to be conserved. The process utilizes 3-D-printed templates as master structures from, which polydimethylsiloxane (PDMS) microstencils are molded. The entire process, from concept to completed stencil, requires approximately one day; however, the majority of this time is budgeted for PDMS curing cycles, so it requires only ∼1-2 person hours to complete. These microstencils were used to pattern metal traces ∼160-1000-µm wide, on three commonly used BioMEMS substrate materialsto pattern rat cortical neuronal cell cultures with radii between ∼300 and 1000 µm-and to pattern organic materials. With the advancement of 3-D-printing technologies, we anticipate that our presented processes will improve in resolution and gain a greater cost advantage over traditional microstencilling methods.[2014-0019]