Human Lung Spheroids as In Vitro Niches of Lung Progenitor Cells With Distinctive Paracrine and Plasticity Properties

Isotta Chimenti, Francesca Pagano, Francesco Angelini, Camilla Siciliano, Giorgio Mangino, Vittorio Picchio, Elena De Falco, Mariangela Peruzzi, Roberto Carnevale, Mohsen Ibrahim, Giuseppe Biondi-Zoccai, Elisa Messina, Giacomo Frati

Research output: Contribution to journalArticle

Abstract

: Basic and translational research on lung biology has discovered multiple progenitor cell types, specialized or facultative, responsible for turnover, renewal, and repair. Isolation of populations of resident lung progenitor cells (LPCs) has been described by multiple protocols, and some have been successfully applied to healthy human lung tissue. We aimed at understanding how different cell culture conditions may affect, in vitro, the phenotype of LPCs to create an ideal niche-like microenvironment. The influence of different substrates (i.e., fibronectin, gelatin, laminin) and the impact of a three-dimensional/two-dimensional (3D/2D) culture switch on the biology of LPCs isolated as lung spheroids (LSs) from normal adult human lung biopsy specimens were investigated. We applied a spheroid culture system as the selective/inductive step for progenitor cell culture, as described in many biological systems. The data showed a niche-like proepithelial microenvironment inside the LS, highly sensitive to the 3D culture system and significantly affecting the phenotype of adult LPCs more than culture substrate. LSs favor epithelial phenotypes and LPC maintenance and contain cells more responsive to specific commitment stimuli than 2D monolayer cultures, while secreting a distinctive set of paracrine factors. We have shown for the first time, to our knowledge, how culture as 3D LSs can affect LPC epithelial phenotype and produce strong paracrine signals with a distinctive secretomic profile compared with 2D monolayer conditions. These findings suggest novel approaches to maintain ex vivo LPCs for basic and translational studies.

SIGNIFICANCE: A lung niche-like microenvironment was created ex vivo inside the pneumosphere. Pneumospheres have a distinctive secretomic profile compared with two-dimensional culture. Three-dimensional culture switch significantly affects lung progenitor phenotype. Lung progenitors in pneumospheres were more committed toward epithelial lineages.

Original languageEnglish
JournalStem cells translational medicine
DOIs
Publication statusE-pub ahead of print - Sep 22 2016

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Stem Cells
Lung
Phenotype
In Vitro Techniques
Cell Culture Techniques
Translational Medical Research
Laminin
Gelatin
Fibronectins
Maintenance

Keywords

  • Journal Article

Cite this

Human Lung Spheroids as In Vitro Niches of Lung Progenitor Cells With Distinctive Paracrine and Plasticity Properties. / Chimenti, Isotta; Pagano, Francesca; Angelini, Francesco; Siciliano, Camilla; Mangino, Giorgio; Picchio, Vittorio; De Falco, Elena; Peruzzi, Mariangela; Carnevale, Roberto; Ibrahim, Mohsen; Biondi-Zoccai, Giuseppe; Messina, Elisa; Frati, Giacomo.

In: Stem cells translational medicine, 22.09.2016.

Research output: Contribution to journalArticle

Chimenti, I, Pagano, F, Angelini, F, Siciliano, C, Mangino, G, Picchio, V, De Falco, E, Peruzzi, M, Carnevale, R, Ibrahim, M, Biondi-Zoccai, G, Messina, E & Frati, G 2016, 'Human Lung Spheroids as In Vitro Niches of Lung Progenitor Cells With Distinctive Paracrine and Plasticity Properties', Stem cells translational medicine. https://doi.org/10.5966/sctm.2015-0374
Chimenti, Isotta ; Pagano, Francesca ; Angelini, Francesco ; Siciliano, Camilla ; Mangino, Giorgio ; Picchio, Vittorio ; De Falco, Elena ; Peruzzi, Mariangela ; Carnevale, Roberto ; Ibrahim, Mohsen ; Biondi-Zoccai, Giuseppe ; Messina, Elisa ; Frati, Giacomo. / Human Lung Spheroids as In Vitro Niches of Lung Progenitor Cells With Distinctive Paracrine and Plasticity Properties. In: Stem cells translational medicine. 2016.
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AU - Angelini, Francesco

AU - Siciliano, Camilla

AU - Mangino, Giorgio

AU - Picchio, Vittorio

AU - De Falco, Elena

AU - Peruzzi, Mariangela

AU - Carnevale, Roberto

AU - Ibrahim, Mohsen

AU - Biondi-Zoccai, Giuseppe

AU - Messina, Elisa

AU - Frati, Giacomo

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N2 - : Basic and translational research on lung biology has discovered multiple progenitor cell types, specialized or facultative, responsible for turnover, renewal, and repair. Isolation of populations of resident lung progenitor cells (LPCs) has been described by multiple protocols, and some have been successfully applied to healthy human lung tissue. We aimed at understanding how different cell culture conditions may affect, in vitro, the phenotype of LPCs to create an ideal niche-like microenvironment. The influence of different substrates (i.e., fibronectin, gelatin, laminin) and the impact of a three-dimensional/two-dimensional (3D/2D) culture switch on the biology of LPCs isolated as lung spheroids (LSs) from normal adult human lung biopsy specimens were investigated. We applied a spheroid culture system as the selective/inductive step for progenitor cell culture, as described in many biological systems. The data showed a niche-like proepithelial microenvironment inside the LS, highly sensitive to the 3D culture system and significantly affecting the phenotype of adult LPCs more than culture substrate. LSs favor epithelial phenotypes and LPC maintenance and contain cells more responsive to specific commitment stimuli than 2D monolayer cultures, while secreting a distinctive set of paracrine factors. We have shown for the first time, to our knowledge, how culture as 3D LSs can affect LPC epithelial phenotype and produce strong paracrine signals with a distinctive secretomic profile compared with 2D monolayer conditions. These findings suggest novel approaches to maintain ex vivo LPCs for basic and translational studies.SIGNIFICANCE: A lung niche-like microenvironment was created ex vivo inside the pneumosphere. Pneumospheres have a distinctive secretomic profile compared with two-dimensional culture. Three-dimensional culture switch significantly affects lung progenitor phenotype. Lung progenitors in pneumospheres were more committed toward epithelial lineages.

AB - : Basic and translational research on lung biology has discovered multiple progenitor cell types, specialized or facultative, responsible for turnover, renewal, and repair. Isolation of populations of resident lung progenitor cells (LPCs) has been described by multiple protocols, and some have been successfully applied to healthy human lung tissue. We aimed at understanding how different cell culture conditions may affect, in vitro, the phenotype of LPCs to create an ideal niche-like microenvironment. The influence of different substrates (i.e., fibronectin, gelatin, laminin) and the impact of a three-dimensional/two-dimensional (3D/2D) culture switch on the biology of LPCs isolated as lung spheroids (LSs) from normal adult human lung biopsy specimens were investigated. We applied a spheroid culture system as the selective/inductive step for progenitor cell culture, as described in many biological systems. The data showed a niche-like proepithelial microenvironment inside the LS, highly sensitive to the 3D culture system and significantly affecting the phenotype of adult LPCs more than culture substrate. LSs favor epithelial phenotypes and LPC maintenance and contain cells more responsive to specific commitment stimuli than 2D monolayer cultures, while secreting a distinctive set of paracrine factors. We have shown for the first time, to our knowledge, how culture as 3D LSs can affect LPC epithelial phenotype and produce strong paracrine signals with a distinctive secretomic profile compared with 2D monolayer conditions. These findings suggest novel approaches to maintain ex vivo LPCs for basic and translational studies.SIGNIFICANCE: A lung niche-like microenvironment was created ex vivo inside the pneumosphere. Pneumospheres have a distinctive secretomic profile compared with two-dimensional culture. Three-dimensional culture switch significantly affects lung progenitor phenotype. Lung progenitors in pneumospheres were more committed toward epithelial lineages.

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