Nanoscale level gelatin-based scaffolds enhance colony formation of porcine testicular germ cells

Abstract

The extracellular matrix is important in cell growth, proliferation, and differentiation. Gelatin, a support for adhering cells, is used for coating culture plate surfaces of several primary and stem cells. However, gelatin characteristics on culture plates and its cell interactions are not understood. Here, we aimed to identify the effect of gelatin topography on culture plates on the proliferation and colony formation of porcine spermatogonial germ cells (pSGC). To generate different surface topographies, gelatin powder was dissolved in H₂O at varying melting temperatures (40, 60, 80, and 120 °C) and coated on the surface of the culture plates. At 40 °C, the pores of the gelatin scaffold were regular ellipses 5-6 μm in diameter and 10-30 nm in thickness. However, at 120 °C, irregular pores 20-30 μm in diameter and 10-20 nm in thickness were obtained. Additionally, the number of attached cells and pSGC colonies were significantly more at 40 °C than at 120 °C after a week of culture. Interestingly, the feeder cells did not settle properly at 120 °C but detached easily from the culture dishes. PSGC colonies were 100 μm in diameter at 40 °C, with small and detached colonies observed at 120 °C. Thus, optimal topography of gelatin was obtained at 40 °C, which was sufficient for the proliferation of feeder cells and the formation of pSGC colonies. Thus, gelatin scaffold conditions at 40 °C and 60 °C were optimal for the derivation and culture of pSGC, and gelatin surface morphology is important for the maintenance of supportive feeder cells for pSGC proliferation and colony formation.

Keywords: Colony formation; Gelatin; Porcine testis cells; Proliferation; Scaffold.