Disease modeling following organoid-based expansion of airway epithelial cells.
Evelien EenjesSander van RietAndre A KroonAnnelies M SlatsP Padmini S J KhedoeAnne Boerema-de MunckMarjon Buscop-van KempenDennis K NinaberIrwin K M ReissHans CleversRobbert J RottierPieter S HiemstraPublished in: American journal of physiology. Lung cellular and molecular physiology (2021)
Air-liquid interface (ALI) cultures are frequently used in lung research but require substantial cell numbers that cannot readily be obtained from patients. We explored whether organoid expansion [three-dimensional (3D)] can be used to establish ALI cultures from clinical samples with low epithelial cell numbers. Airway epithelial cells were obtained from tracheal aspirates (TA) from preterm newborns and from bronchoalveolar lavage (BAL) or bronchial tissue (BT) from adults. TA and BAL cells were 3D-expanded, whereas cells from BT were expanded in 3D and 2D. Following expansion, cells were cultured at ALI to induce differentiation. The impact of cell origin and 2D or 3D expansion was assessed with respect to 1) cellular composition, 2) response to cigarette smoke exposure, and 3) effect of Notch inhibition or IL-13 stimulation on cellular differentiation. We established well-differentiated ALI cultures from all samples. Cellular compositions (basal, ciliated, and goblet cells) were comparable. All 3D-expanded cultures showed a similar stress response following cigarette smoke exposure but differed from the 2D-expanded cultures. Higher peak levels of antioxidant genes HMOX1 and NQO1 and a more rapid return to baseline, and a lower unfolded protein response was observed after cigarette smoke exposure in 3D-derived cultures compared to 2D-derived cultures. In addition, TA- and BAL-derived cultures were less sensitive to modulation by DAPT or IL-13 than BT-derived cultures. Organoid-based expansion of clinical samples with low cell numbers, such as TA from preterm newborns is a valid method and tool to establish ALI cultures.
Keyphrases
- induced apoptosis
- single cell
- cell cycle arrest
- end stage renal disease
- oxidative stress
- pregnant women
- low birth weight
- signaling pathway
- cell therapy
- mesenchymal stem cells
- dna methylation
- chronic kidney disease
- cell proliferation
- gene expression
- gestational age
- preterm infants
- preterm birth
- small molecule
- endoplasmic reticulum stress
- ionic liquid
- newly diagnosed
- quantum dots
- binding protein
- bone marrow
- pi k akt