Human anterior segment perfusion cultures are frequently used for trabecular meshwork research. However, this model requires the use of whole eye globes which are expensive. Here, we describe a method using human corneal rims as an alternative to anterior segments for perfusion culture. In this method, the human corneal rim is glued to a three-dimensional-printed perfusion plates instead of using mechanical compression in traditional perfusion culture setup. The corneal rim and perfusion plate are placed onto a special holder for perfusion culture. This method is affordable and easy to set up. The instruments for traditional perfusion culture studies can be used for this model without modification.