Cryobanking of Human Distal Lung Epithelial Cells for Preservation of Their Phenotypic and Functional Characteristics

Konda, Bindu; Mulay, Apoorva; Yao, Changfu; Beil, Stephen; Huynh, Carissa A.; Tourtellotte, Warren G.; Rampolla, Reinaldo; Chen, Peter C.; Carraro, Gianni; Stripp, Barry R.

doi:10.1165/rcmb.2021-0507ma

Cited by 4 publications

(2 citation statements)

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“…Distal lung preparations from adult donor lung explants from 5 individuals (primary lung 1–5, PL 1–5) were cryopreserved using methods we recently described (ref. 21 and depicted in Figure 1A ). After thawing, AEC2s were purified using FACS to isolate cells coexpressing EPCAM and the AEC2-selective surface marker HTII-280 ( 35 ).…”

Section: Resultsmentioning

confidence: 93%

“…With the advent of 3D culture models, we and others have developed methods for the in vitro culture of human AEC2-like cells derived from 1° fetal ( 7 ), 1° adult ( 10 , 14 – 16 , 21 , 22 ), or induced pluripotent stem cell (iPSC) sources ( 13 , 22 , 23 ). These methods have enabled the in vitro maintenance of functional human AEC2-like cells that share similar transcriptional and ultrastructural properties to their in vivo or freshly isolated 1° AEC2 counterparts, including the capacity to produce surfactant proteins and phospholipids ( 13 – 16 , 23 ).…”

Section: Introductionmentioning

confidence: 99%

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Culture impact on the transcriptomic programs of primary and iPSC-derived human alveolar type 2 cells

Alysandratos¹,

Alba²,

Yao³

et al. 2023

JCI Insight

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Dysfunction of alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of lung alveoli, is implicated in pulmonary disease pathogenesis, highlighting the importance of human in vitro models. However, AEC2-like cells in culture have yet to be directly compared to their in vivo counterparts at single cell resolution. Here, we perform head-to-head comparisons between the transcriptomes of fresh primary (1 o ) adult human AEC2s, their cultured progeny, and human induced pluripotent stem cell-derived AEC2s (iAEC2s). We find each population occupies a distinct transcriptomic space with cultured AEC2s (1 o and iAEC2s) exhibiting similarities to and differences from freshly purified 1 o cells. Across each cell type, we find an inverse relationship between proliferative and maturation states, with pre-culture 1 o AEC2s being most quiescent/mature and iAEC2s being most proliferative/least mature. Cultures of either type of human AEC2 do not generate detectable alveolar type 1 cells in these defined conditions; however, a subset of iAEC2s co-cultured with fibroblasts acquires a "transitional cell state" described in mice and humans to arise during fibrosis or following injury. Hence, we provide direct comparisons of the transcriptomic programs of 1 o and engineered AEC2s, two in vitro models that can be harnessed to study human lung health and disease.

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Section: Resultsmentioning

confidence: 93%