Isolation and Culture of Alveolar Epithelial Type I and Type II Cells from Rat Lungs

Gonzalez, Robert; Dobbs, Leland G.

doi:10.1007/978-1-62703-125-7_10

Cited by 57 publications

(48 citation statements)

References 9 publications

(10 reference statements)

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“…Mouse AT2 cells were isolated via a dispase-based digestion method as previously described (43). Rat type AT2 cells were isolated via an elastase-based digestion method, followed by panning with rat IgG-coated plates, and cultured on 12-well tissue culture plastic as previously described for 2 days prior to stimulation (65). Primary human AT2 cells were isolated from human lungs from deidentified organ donors whose lungs were not suitable for transplantation.…”

Section: Animals Gpr116supporting

confidence: 47%

Epithelial Gpr116 regulates pulmonary alveolar homeostasis via Gq/11 signaling

et al. 2017

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Section: Animals Gpr116supporting

confidence: 47%

Epithelial Gpr116 regulates pulmonary alveolar homeostasis via Gq/11 signaling

et al. 2017

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“…3g,h). Conversely, under conditions that promote differentiation into an AT1 fate (39), addition of recombinant Wnt5a inhibited this transdifferentiation 2.6-fold (21±1% vs 8±1% with Wnt5a) (Fig. 3i).…”

Section: Resultsmentioning

confidence: 97%

“…Under culture conditions that promote maintenance of the AT2 cell phenotype (39), addition of the Wnt antagonist Dickkopf 3 (Dkk3) (40) increased by 3.8-fold the percentage of cells that reprogrammed to AT1 identity (2.5±1.5% vs. 9.5±0.1% with Dkk3) (Fig. 3g,h).…”

Section: Resultsmentioning

confidence: 99%

Single-cell Wnt signaling niches maintain stemness of alveolar type 2 cells

Nabhan

Brownfield

Harbury

et al. 2018

Science

593

696

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Lung alveoli are lined by squamous alveolar epithelial type 1 (AT1) epithelial cells that facilitate gas exchange, and neighboring AT2 cells that synthesize and secrete surfactant. Alveoli are maintained by intermittent activation of rare ‘bifunctional’ AT2 cells that retain surfactant biosynthesis function but also serve as stem cells, generating new AT1 cells and self-renewing throughout adult life. While stem cell proliferation is controlled by EGFR/KRAS signaling, how the stem cells are selected, maintained, and the fates of their daughter cells controlled are unknown. Here we show that expression of the Wnt target gene Axin2 in mouse lung identifies a rare, stable subpopulation of AT2 cells with stem cell activity. Many lie near single fibroblasts that express Wnt5a and other Wnt genes, and genetically targeting Wnt secretion by fibroblasts depletes the Axin2+ AT2 stem cell population. Axin2 turns off when daughter cells leave the Wnt niche and transdifferentiate into AT1 cells, and sustaining Wnt signaling blocks transdifferentiation whereas abrogation of Wnt signaling promotes it, both in vivo and in vitro. Upon severe alveolar epithelial injury, Axin2 is induced throughout the AT2 population, recruiting ‘ancillary’ AT2 cells into a progenitor role. Niche expression of Wnt5a and the Wnt secretion mediator Porcupine is unchanged by injury, but Wnt7b and several other Wnt genes are broadly induced along with Porcupine in AT2 cells, and pharmacologic or genetic inhibition of this autocrine Wnt signaling impairs the AT2 proliferative response. The results support a model in which individual AT2 cells reside in single cell fibroblast niches that provide a short-range paracrine (or "juxtacrine") Wnt signal that selects and maintains alveolar stem cell identity and proliferative capacity, while severe injury induces AT2 autocrine Wnt signals that transiently expand the stem cell pool during repair.

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“…The evolution of ATII cell culture methods from various mammalian species is given in Table 2. Compared to ATII cells, there are fewer studies of ATI cells, but these cells can be selectively isolated and cultured, as described in detail for rats by Gonzalez and Dobbs (2013).…”

Section: Modern Alveolar Epithelial Cell Culturesmentioning

confidence: 44%