Coupling 3D Printing and Novel Replica Molding for In House Fabrication of Skeletal Muscle Tissue Engineering Devices

Abstract

The transition from 2D to 3D engineered tissue cultures is changing the way biologists can perform in vitro functional studies. However, there has been a paucity in the establishment of methods required for the generation of microdevices and cost‐effective scaling up. To date, approaches including multistep photolithography, milling and 3D printing have been used that involve specialized and expensive equipment or time‐consuming steps with variable success. Here, a fabrication pipeline is presented based on affordable off‐the‐shelf 3D printers and novel replica molding strategies for rapid and easy in‐house production of hundreds of 3D culture devices per day, with customizable size and geometry. This pipeline is applied to generate tissue engineered skeletal muscles in vitro using human induced pluripotent stem cell‐derived myogenic progenitors. These production methods can be employed in any standard biomedical laboratory.

Hooked on mushrooms: Preparation and mechanics of a bioinspired soft probabilistic fastener

“…This means that if the 3D printed structure is not completely cured, it will react with the uncured ecoflex and it will chemically bond to it, making the peeling off impossible. Similar considerations are relevant when casting PDMS on ecoflex [26]. If the latter is not properly cured, the two silicone based elastomers will react and seal together.…”

“…Attachment and subsequent detachment from an opposing surface such as a fabric can break either the small 3D printed features or damage the fabric itself. Therefore, we replicated the 3D printed structure in polydimethylsiloxane elastomer (PDMS) using a double molding process [21][22][23]. At first, the positive 3D printed structure was replicated as negative in an elastomer ecoflex 0030.…”

The Field Guide to 3D Printing in Microscopy

“…Additionally, by bonding printed channels to transparent PMMA sheets, it is possible to produce highly complex arrays, such as straight, spiral, serpentine, curvilinear, and contraction-expansion (70). In some cases, designs such as T-shape pillars are not possible to obtain via other traditional fabrication methods in a single demolding step, but only by using commercial 3D printers (71).…”

“…Once the 3D cell cultures were established in the chamber, a suspension of hiPSC-derived myogenic progenitors and biocompatible hydrogels made of fibrin and Matrigel (74) was introduced. Differentiation resulted in the successful formation of a TESM culture with a comparable composition to other cultures, including myofibers (71). In addition, organ-on-chip managed to mimic liver interstitial structures containing endothelial cells and primary hepatocytes (75), produce a drug screening assay to assess cell reaction (76), and test for injury (77), hepatitis B infections and even alcohol-driven injury (78).…”

“…Thanks to this ingenious method, those printers are equally or even more inexpensive than) FDM printers, and, as they cure one complete layer at a time without the need of moving a laser (or a printhead), they are also faster than SLA or FDM. The inexpensive availability of those printers, together with their speed and precision may open the possibility of using mSLA printers in the lab (71). Clear/transparent materials are achievable with liquid resins.…”