Objectives: In a tissue engineering field, co-culture techniques of different kind have been widely researched in recent years to form physiologically relevant cell structures for different tissue reconstructions. The treatment methods of that kind are promising opportunity to be used also in the treatment of corneal blindness, which is still mainly treated with traditional tissue graft from a donor. Tissue grafts cannot still be used to treat one of the most common cause behind the corneal blindness -limbal stem cell deficiency (LSCD), due the lack of host stem cells. Thus, alternative opportunities are needed. Objectives of this research were to find a suitable medium for human adipose stem cells (hASCs), from those that had already been shown to be suitable for human corneal epithelial cells (hCECs), and use it to co-culture hydrogel- embedded hASCs with hCECs. Results will possibly help us to evaluate the functionality of this kind of 3D structure better as a treatment method of the corneal damages and LSCD.
Materials and methods: HASCs were first cultured in three different mediums to find a suitable one to use in co-culturing. Following 2D co-culturing occurred within indirect- and direct contact between the immortalized hCECs and hASCs. Research was accomplished by culturing the hASCs inside the hyaluronic acid- hydrogel. Cells' behaviour during these experiments was evaluated with phase-contrast microscope, proliferation- and live/dead assays and immunofluorescent stainings.
Results and conclusions: HASCs grew well in tested co-culture medium and appeared to have enhancing effect on the growth and differentiation of the co-cultured immortalized hCECs. When cultured in 3D hydrogel structure, hASCs were able to survive and proliferate within right cell densities. These results indicate that 3D co-cultures with hCECs and hydrogel- embedded hASCs might have potential for future applications in ocular surface reconstruction
