Abstract
Development of a simple, straightforward 3D fabrication method to culture cells in 3D, without relying on any complex fabrication methods, remains a challenge. In this paper, we describe a new technique that allows fabrication of scalable 3D cell-laden hydrogel structures easily, without complex machinery: the technique can be done using only apparatus already available in a typical cell biology laboratory. The fabrication method involves micro dip-coating of cell-laden hydrogels covering the surface of a metal bar, into the cross-linking reagents calcium chloride or barium chloride to form hollow tubular structures. This method can be used to form single layers with thickness ranging from 126 to 220 µm or multilayered tubular structures. This fabrication method uses alginate hydrogel as the primary biomaterial and a secondary biomaterial can be added depending on the desired application. We demonstrate the feasibility of this method, with survival rate over 75% immediately after fabrication and normal responsiveness of cells within these tubular structures using mouse dermal embryonic fibroblast cells and human embryonic kidney 293 cells containing a tetracycline-responsive, red fluorescent protein (tHEK cells).
Highlights
Conventional 2D, monolayer cell culture still remains the main approach for the study of cell biology, regenerative medicine, and drug discovery (Gies et al, 2002)
Our strategy for producing cell-laden tubular structures was to dip a stainless steel rod into a suspension of cells in an alginatebased buffer and to transfer the rod, still coated with a thin layer of this suspension, into a solution of divalent cations that would cross-link the alginate into a hydrogel (Figure 1A)
We have developed a new rapid 3D biofabrication technique for making alginate hydrogel tubular structures
Summary
Conventional 2D, monolayer cell culture still remains the main approach for the study of cell biology, regenerative medicine, and drug discovery (Gies et al, 2002). The main barrier in conventional scaffold-based tissue engineering approaches is the inability to position living cells precisely to mimic 3D tissue. Repopulation of decellularized tissues and organs has been reported to regenerate 3D tissue and can be used as a platform for drug discovery and organ transplantation, but this approach relies. There is, a significant problem with current 3D biofabrication approaches: they can generate simple and precise 3D structures, they rely on specialized bioprinting machinery that is not accessible to many cell biologists. A simple, controllable, and straightforward 3D biofabrication method that does not involve complicated machinery such as bioprinting platforms would be very valuable and effective to create biomimetic 3D structures
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