TY - JOUR
T1 - Disease modeling following organoid-based expansion of airway epithelial cells
AU - Eenjes, Evelien
AU - van Riet, Sander
AU - Kroon, Andre A
AU - Slats, Annelies M
AU - Khedoe, P Padmini S J
AU - Boerema-de Munck, Anne
AU - Buscop-van Kempen, Marjon
AU - Ninaber, Dennis K
AU - Reiss, Irwin K M
AU - Clevers, Hans
AU - Rottier, Robbert J
AU - Hiemstra, Pieter S
PY - 2021/10/1
Y1 - 2021/10/1
N2 - 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.
AB - 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.
KW - Adult
KW - Bronchi/cytology
KW - Bronchoalveolar Lavage Fluid/cytology
KW - Cell Culture Techniques
KW - Cell Differentiation/physiology
KW - Cells, Cultured
KW - Epithelial Cells/cytology
KW - Heme Oxygenase-1/metabolism
KW - Humans
KW - Infant, Newborn
KW - Interleukin-13/metabolism
KW - NAD(P)H Dehydrogenase (Quinone)/metabolism
KW - Organoids/cytology
KW - Receptors, Notch/antagonists & inhibitors
KW - Respiratory Mucosa/cytology
KW - Smoke/adverse effects
KW - Tobacco Products/adverse effects
KW - Unfolded Protein Response/drug effects
U2 - 10.1152/ajplung.00234.2020
DO - 10.1152/ajplung.00234.2020
M3 - Article
C2 - 34378410
SN - 1040-0605
VL - 321
SP - L775-L786
JO - American journal of physiology. Lung cellular and molecular physiology
JF - American journal of physiology. Lung cellular and molecular physiology
IS - 4
ER -