3D Cultures of Salivary Gland Cells in Native or Gelled Egg Yolk Plasma, Combined with Egg White and 3D-Printing of Gelled Egg Yolk Plasma

• Main Authors: André M. Charbonneau, Joseph M. Kinsella, Simon D. Tran
• Format: Article
• Language: English
• Published: MDPI AG 2019-10-01
• Series: Materials
• Subjects: 3d-bioprinting; 3d-printing; salivary glands; 3d-cryo well insert; histology; egg yolk plasma; egg white; gel; ki-67; rheology; cell culture; tissue engineering
• Online Access: https://www.mdpi.com/1996-1944/12/21/3480

For salivary gland (SG) tissue engineering, we cultured acinar NS-SV-AC cell line or primary SG fibroblasts for 14 days in avian egg yolk plasma (EYP). Media or egg white (EW) supplemented the cultures as they grew in 3D-Cryo histology well inserts. In the second half of this manuscript, we measured EYP’s freeze-thaw gelation and freeze-thaw induced gelled EYP (_GEYP), and designed and tested further _GEYP tissue engineering applications. With a 3D-Cryo well insert, we tested _GEYP as a structural support for 3D cell culture or as a bio-ink for 3D-Bioprinting fluorescent cells. In non-printed EYP + EW or _GEYP + EW cultures, sagittal sections of the cultures showed cells remaining above the well’s base. Ki-67 expression was lacking for fibroblasts, contrasting NS-SV-AC’s constant expression. Rheological viscoelastic measurements of _GEYP at 37 °C on seven different freezing periods showed constant increase from 0 in mean storage and loss moduli, to 320 Pa and 120 Pa, respectively, after 30 days. We successfully 3D-printed _GEYP with controlled geometries. We manually extruded _GEYP bio-ink with fluorescence cells into a 3D-Cryo well insert and showed cell positioning. The 3D-Cryo well inserts reveal information on cells in EYP and we demonstrated _GEYP cell culture and 3D-printing applications.