3D micro-organisation printing of mammalian cells to generate biological tissues

Abstract

Significant strides have been made in the development of in vitro systems for disease modelling. However, the requirement of microenvironment control has placed limitations on the generation of relevant models. Herein, we present a biological tissue printing approach that employs open-volume microfluidics to position individual cells in complex 2D and 3D patterns, as well as in single cell arrays. The variety of bioprinted cell types employed, including skin epithelial (HaCaT), skin cancer (A431), liver cancer (Hep G2), and fibroblast (3T3-J2) cells, all of which exhibited excellent viability and survivability, allowing printed structures to rapidly develop into confluent tissues. To demonstrate a simple 2D oncology model, A431 and HaCaT cells were printed and grown into tissues. Furthermore, a basic skin model was established to probe drug response. 3D tissue formation was demonstrated by co-printing Hep G2 and 3T3-J2 cells onto an established fibroblast layer, the functionality of which was probed by measuring albumin production, and was found to be higher in comparison to both 2D and monoculture approaches. Bioprinting of primary cells was tested using acutely isolated primary rat dorsal root ganglia neurons, which survived and established processes. The presented technique offers a novel open-volume microfluidics approach to bioprint cells for the generation of biological tissues.

Review of Low-Cost 3D Bioprinters: State of the Market and Observed Future Trends

“…For its printing technology, the Biopixlar uses a "microfluidic hydrodynamic confined flow" technology for building biological tissues from individual cells. [213][214][215] Specifically, the printer nozzle, in this case, is essentially a microfluidic device whose tip has one outflow channel in the center and two inflow ones at the sides (Fig. 7b).…”

“…As shown in a recent publication, 214 the Biopixlar was able to print up to three different cell types for fabricating 2D/3D complex tissue structures at a single-cell resolution. The printing process was shown to have maintained ~96-97% cell viability at 2 h postfabrication and a higher than 99% cell survivability after 24 h, when skin cancer (A431) and epithelial (HaCaT) cell types were used.…”

Nanomedicine for Gene Delivery and Drug Repurposing in the Treatment of Muscular Dystrophies

“…In this context, 3D culture systems have gained increasing interest as they are able to provide accurate models of organs or tissue physiology and associated disorders. However, they are not exempt from limitations, such as lack of nutrients and oxygen distribution, and accumulation of wastes into the core of 3D culture [ 236 , 238 , 239 ]. Diverse biomimetic engineered muscle constructs, such as scaffold or organoid cell culture, have demonstrated structural and functional characteristics similar to native muscle, mimicking the tissue complexity [ 240 , 241 , 242 ].…”

Dorsal Root Injury—A Model for Exploring Pathophysiology and Therapeutic Strategies in Spinal Cord Injury

“…3D bioprinting has rapidly developed as a highly attractive technology for b metic implants that is able to support neuronal survival, counteract disease driving cesses and support functional repair following spinal cord injury [144]. Based on det information on the underlying pathophysiological processes and conditions of the pa 3D bioprinted implants may be designed to match patient-specific conditions [165] cent studies have demonstrated 3D bioprinted constructs that support neurite outgr from dorsal root ganglion neurons in vitro [166][167][168]. Dorsal root injury would be a u in vivo model for exploring these developments and their further translation to the in spinal cord.…”

“…Based on detailed information on the underlying pathophysiological processes and conditions of the patient, 3D bioprinted implants may be designed to match patient-specific conditions [ 165 ]. Recent studies have demonstrated 3D bioprinted constructs that support neurite outgrowth from dorsal root ganglion neurons in vitro [ 166 , 167 , 168 ]. Dorsal root injury would be a useful in vivo model for exploring these developments and their further translation to the injured spinal cord.…”