TUNICELL for 3D cultivation of human dermal fibroblasts

Ingrid Weløy Aarseth has just finished her Master of Science in Biotechnology at the Norwegian University of Science and Technology (NTNU). In her thesis she worked with functionalized alginate-nanocellulose hydrogels for 3D cultivation of human dermal fibroblasts. Ingrid used TUNICELL TTC, a transparent biomaterial well-suited for imaging, to bioengineer 3D constructs that were seeded with cells.

Photo: Geir Moen
Photo: Geir Moen

Human dermal fibroblasts are important members of the connective-tissue cell family. They are often used in tissue engineering as they are easy to grow in culture, and have high rates of activity in wound healing. In the body, fibroblasts migrate to injured tissue sites to produce a collagenous matrix to heal the injured tissue.

Processed CLSM images of NHDF cells seeded on crosslinked hydrogel disks. Labels on the left indicate hydrogels formulated with RGD coupled alginate (2.25% w/v) (RGD) or the negative control oxidized alginate (2.25% w/v) (Ox) and TUNICELL TTC. Labels at the top indicate the type of alginate used (LHS-3 or Pro22 at 0.75% w/v) and the TTC+M/alginate ratios (70/30 or 80/20). Cells were stained with Phalloidin 568 and DAPI to show cytoskeleton (red) and nucleus (blue) after incubation for 48 hours. Scale bars = 100 μm.

This novel approach using bioprinting in combination with a three dimensional culture system provides benefits to studying cell and tissue function since these cells behave much more like they do in the body. Human dermal fibroblasts resemble their in vivo morphology, with elongated and spindle-shaped morphology when in 3D, compared to 2D where they have fan-shaped lamellae and slower proliferation.

Processed CLSM images of NHDF cells seeded on bioprinted hydrogels composed of LHS-3-7030RGD. A, B, C and D refer to different imaged replicates. Cells were stained with Phalloidin 488 and DAPI to show the cytoskeleton (red) and nucleus (blue) after incubation for 14 days. Scale bars = 100 μm.

Creating better in vitro microenvironments for cells that are more physiologically representative to the body is important in order to establish tissue models that can be used in cancer and drug research.

Read Ingrid Weløy Aarseth’s full thesis published in the NTNU database here: https://ntnuopen.ntnu.no/ntnu-xmlui/handle/11250/3012761

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