Celkova Patricia, Seydoux Emilie, De Groof Susan, Müller Loretta
Division of Pediatric Respiratory Medicine and Allergology, Department of Pediatrics, Inselspital, Bern University Hospital, 3010 Bern, Switzerland.
Lung Precision Medicine (LPM), Department for BioMedical Research (DBMR), University of Bern, 3008 Bern, Switzerland.
Life (Basel). 2025 Jun 14;15(6):958. doi: 10.3390/life15060958.
Cultures of primary human nasal epithelial cells (hNECs) differentiated at the air-liquid interface (ALI) represent a sophisticated and widely used model of the human upper respiratory epithelium. Despite the availability of various cell culture insert types and the well-established understanding that different culture media influence the cell culture characteristics, the possible impact of the insert brand remains rather underexplored. We cultured hNECs from nineteen healthy adult donors on three distinct brands of commercially available inserts-Corning Transwell, CELLTREAT, and ThinCert-and compared the ciliary activity and cellular composition of the cultures using high-speed video microscopy and flow cytometry, respectively. Additionally, we employed an alternative method of hNEC culture setup-the inverted condition-wherein the hNECs were seeded on the basal side of the insert with the idea to avoid mucus accumulation. Our results show that ciliary activity and cell type composition did not differ between insert types for both culture conditions. However, we found a higher ciliary beat frequency and a lower active (ciliated) area in the inverted setup compared to the conventional setup across all three insert brands. These findings indicate that all three mentioned insert types yield comparable cell cultures.
在气液界面(ALI)分化的原代人鼻上皮细胞(hNECs)培养物代表了一种复杂且广泛使用的人上呼吸道上皮模型。尽管有各种细胞培养插入物类型可供选择,并且人们也充分认识到不同的培养基会影响细胞培养特性,但插入物品牌的潜在影响仍未得到充分探索。我们将来自19名健康成年供体的hNECs培养在三种不同品牌的市售插入物上——康宁Transwell、CELLTREAT和ThinCert——并分别使用高速视频显微镜和流式细胞术比较了培养物的纤毛活性和细胞组成。此外,我们采用了一种hNEC培养设置的替代方法——倒置条件,即将hNECs接种在插入物的基底侧,以避免黏液积聚。我们的结果表明,两种培养条件下,不同插入物类型之间的纤毛活性和细胞类型组成没有差异。然而,我们发现,与传统设置相比,在所有三个插入物品牌中,倒置设置下的纤毛摆动频率更高,活跃(有纤毛)面积更小。这些发现表明,上述三种插入物类型都能产生可比的细胞培养物。