Automating hESC differentiation with 3D printing and legacy liquid handling solutions.

Historically, the routine use of laboratory automation solutions has been prohibitively expensive for many laboratories. As legacy hardware has begun to emerge on the secondary market, automation is becoming an increasingly affordable option to augment workflow in virtually any laboratory. To assess the utility of legacy liquid handling in stem cell differentiation, a used liquid handling robot was purchased at auction to automate a stem cell differentiation protocol that gives rise to CD14 + CD45+ mononuclear cells. To maintain sterility, the automated liquid handling robot was housed in a custom constructed HEPA filtered enclosure. A custom cell scraper and a disposable filter box were designed and 3D printed to permit the robot intricate cell culture actions required by the protocol. All files for the 3D printed labware are uploaded and are freely available. •A used liquid handling robot was used to automate an hESC to monocyte differentiation protocol.•The robot-performed protocol induced monocytes as effectively as human technicians.•Custom 3D printed labware was made to permit certain cell culture actions and are uploaded for free access.