Tissue engineering is emerging as a modern medicine fascination towards the establishment of human tissue banks; yet, these approaches typically involve cultures of only one type of cell and, therefore, do not recapitulate the native tissue physiology in toto. Co-culture models, comprised of different cell types, can potentially create the next level of complexity. However, conventional approaches involving multiple cell types and cell culture inserts do have limitations. To this end, here we demonstrate a novel cell culture insert that allows the use of any custom-made scaffold, free-flow of fluids/gases, dual cell seeding on either sides of the insert, easy stacking of multiple inserts and resizing it to any multi well plate format as well as culture dishes. To prove the concept, electrospun silk fibroin scaffold was clipped onto the insert and was used for co-culturing of keratinocytes and fibroblast cells. The results indicated a successful fabrication of spatially organized skin tissue constructs having epidermal and dermal equivalent histology. Cell-laden inserts were stacked and used for simulated transportation studies. However, he conditions need further fine-tuning. All together, the results indicated that the novel cell culture insert with silk fibroin scaffold could be used as a facile, versatile and scalable approach to fabricate and transport 3D co-cultured tissue constructs in vitro, including but not limited to skin. The resultant tissue constructs can be explored for therapeutic applications, for instance as artificial skin substitute in wound healing, and for toxicological applications, for instance as reconstructed skin tissue model in skin irritation testing.
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Tissue engineering is emerging as a modern medicine fascination towards the establishment of human tissue banks; yet, these approaches typically involve cultures of only one type of cell and, therefore, do not recapitulate the native tissue physiology in toto. Co-culture models, comprised of different cell types, can potentially create the next level of complexity. However, conventional approaches involving multiple cell types and cell culture inserts do have limitations. To this end, here we demonstrate a novel cell culture insert that allows the use of any custom-made scaffold, free-flow of fluids/gases, dual cell seeding on either sides of the insert, easy stacking of multiple inserts and resizing it to any multi well plate format as well as culture dishes. To prove the concept, electrospun silk fibroin scaffold was clipped onto the insert and was used for co-culturing of keratinocytes and fibroblast cells. The results indicated a successful fabrication of spatially organized skin tissue constructs having epidermal and dermal equivalent histology. Cell-laden inserts were stacked and used for simulated transportation studies. However, he conditions need further fine-tuning. All together, the results indicated that the novel cell culture insert with silk fibroin scaffold could be used as a facile, versatile and scalable approach to fabricate and transport 3D co-cultured tissue constructs in vitro, including but not limited to skin. The resultant tissue constructs can be explored for therapeutic applications, for instance as artificial skin substitute in wound healing, and for toxicological applications, for instance as reconstructed skin tissue model in skin irritation testing.
