Simple, low-cost and well-performing method for isolation of epithelial cells from nasal polyps with outgrowth technique

Background Epithelial cells (EC) are an important component of the pathomechanism of type 2 inflammation in chronic rhinosinusitis with nasal polyps. Therefore, it is important to establish a robust method for isolation and culture of EC from nasal polyps to enable further investigation. In this study, we evaluated the feasibility of the outgrowth technique to isolate EC from nasal polyps.

Methods The outgrowth technique was performed to isolate EC from nasal polyps (n = 3). Proliferation of isolated EC was evaluated in cell culture until the 3rd passage. Flow cytometry with anti-cytokeratin, anti-p63, and anti-Ki-67 was used to identify EC and evaluate their differentiation and proliferation. A functional test was performed by determining relevant proteins of type 2 inflammation by ELISA, representative interleukin-33 and periostin.

Results Using outgrowth technique, EC could be isolated from all tissue samples. Isolated EC showed a proliferation rate of approximately 7- to 23-fold (13.48 ± 1.753) until the 3rd passage every 6 days. Flow cytometry showed that over 97% of the isolated cells (97.02 ± 0.2194%) were cytokeratin and p63 positive and over 86% of them (86.45 ± 2.532%) were Ki-67 positive. Interleukin-33 and periostin were detectable in the supernatant.

Conclusions We introduce a simple, low-cost, and well-performing method for isolating EC from nasal polyps with the outgrowth technique.

Publication History

Article published online:
12 May 2023