Expansion of Primary Bronchial Epithelial Cell Cultures

Margarit, G.; Belda, J.; Casan, P.; Sanchis, J.

doi:10.1016/S1579-2129(06)60273-5

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The aim of this study was to develop a way to increase the production of human bronchial epithelial cells (BEC) in primary cultures. Materialand methodsA total of 12 samples (9 from surgical specimens and 3 from endoscopic biopsies) were processed on plates coated with type I collagen with growth medium supplemented for BEC. When cell proliferation started, the explants were removed for successive subculturing. The remaining cells were left to proliferate and were trypsinized after 50% confluence. We recorded the number of cells obtained, cell viability, and the percentage positive for cytokeratin 7. ResultsThe total number of cells obtained by this method was 3-fold the number of human BEC obtained with simple primary cultures. The maximum number of subcultures was 5, mean (SD) cell viability was 91.9% (11.7%), and the percentage of cells positive for cytokeratin 7 was 30.71% (10.68%). ConclusionsThe described method for expanding primary BEC cultures increases cell production." ] "es" => array:1 [ "resumen" => "ObjetivoLOS cultivos celulares son un buen modelo para el estudio de las enfermedades pulmonares, pero son difíciles de reproducir y producen un número limitado de células. El objetivo de este estudio ha sido desarrollar un método que incrementase la producción de células epiteliales bronquiales (CEB) humanas en cultivos primarios. Materialy métodosSe procesó un total de 12 muestras (9 procedentes de muestras quirúrgicas y 3 de biopsias endoscópicas) en placas recubiertas de colágeno tipo I con medio suplementado para CEB. Al iniciarse la proliferación celular a su alrededor, los explantes se extrajeron y subcultivaron sucesivamente. Las células restantes se dejaron proliferar y se tripsinizaron tras alcanzar más del 50% de confluencia. Se valoraron el número de células obtenidas, la viabilidad y la citoqueratina 7. ResultadosEl número total de células obtenidas con este método superó en una media de 3 veces el número de CEB humanas obtenidas en cultivos primarios simples. El número máximo de subcultivos fue de 5, la viabilidad media (± desviación estándar) fue de 91,9 ± 11,7% y el porcentaje de células positivas para la citoqueratina 7 del 30,71 ± 10,68%. 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"volumen" => "4" "paginaInicial" => "210" "paginaFinal" => "217" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/2044738" "web" => "Medline" ] ] ] ] ] ] ] ] 4 => array:3 [ "identificador" => "bib5" "etiqueta" => "5" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Growth and differentiation of conducting airway epithelial cells in culture" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "R Wu" 1 => "YH Zhao" 2 => "MMJ Chang" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Eur Respir J" "fecha" => "1997" "volumen" => "10" "paginaInicial" => "2398" "paginaFinal" => "2403" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/9387971" "web" => "Medline" ] ] ] ] ] ] ] ] 5 => array:3 [ "identificador" => "bib6" "etiqueta" => "6" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "OVTL 12/30 (keratin 7 antibody) is a marker of glandular differentiation in lung cancer" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "FJ van der Molengraft" 1 => "CC van Niekerk" 2 => "PH Jap" 3 => "LG Poels" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Histopathology" "fecha" => "1993" "volumen" => "22" "paginaInicial" => "35" "paginaFinal" => "38" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/7679658" "web" => "Medline" ] ] ] ] ] ] ] ] 6 => array:3 [ "identificador" => "bib7" "etiqueta" => "7" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Culture and characterisation of human bronchial epithelial cells in vitro" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "RJ Sapsford" 1 => "JH Wang" 2 => "JL Devalia" 3 => "RJ Davies" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" 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Archivos de Bronconeumología

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Vol. 41. Issue 9.

Pages 524-527 (September 2005)

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Pages 524-527 (September 2005)

Techniques and Procedures

DOI: 10.1016/S1579-2129(06)60273-5

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Expansion of Primary Bronchial Epithelial Cell Cultures

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G. Margarit

Corresponding author

gmargarit@hsp.santpau.es

Correspondence: Dra. G. Margarit. Antoni M. Claret, 167. 08025 Barcelona. España

, J. Belda, P. Casan, J. Sanchis

Departamento de Neumología, Hospital de la Santa Creu i de Sant Pau, Facultat de Medicina, Universitat Autònoma de Barcelona, Barcelona, Spain

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Objective

Cell cultures provide a good model for studying lung diseases but they are difficult to reproduce and the number of cells obtained is limited. The aim of this study was to develop a way to increase the production of human bronchial epithelial cells (BEC) in primary cultures.

Materialand methods

A total of 12 samples (9 from surgical specimens and 3 from endoscopic biopsies) were processed on plates coated with type I collagen with growth medium supplemented for BEC. When cell proliferation started, the explants were removed for successive subculturing. The remaining cells were left to proliferate and were trypsinized after 50% confluence. We recorded the number of cells obtained, cell viability, and the percentage positive for cytokeratin 7.

Results

The total number of cells obtained by this method was 3-fold the number of human BEC obtained with simple primary cultures. The maximum number of subcultures was 5, mean (SD) cell viability was 91.9% (11.7%), and the percentage of cells positive for cytokeratin 7 was 30.71% (10.68%).

Conclusions

The described method for expanding primary BEC cultures increases cell production.

Key Words:

Primary cultures

Bronchial epithelial cells

Culture expansion

Explants

Cytokeratin 7

Objetivo

LOS cultivos celulares son un buen modelo para el estudio de las enfermedades pulmonares, pero son difíciles de reproducir y producen un número limitado de células. El objetivo de este estudio ha sido desarrollar un método que incrementase la producción de células epiteliales bronquiales (CEB) humanas en cultivos primarios.

Materialy métodos

Se procesó un total de 12 muestras (9 procedentes de muestras quirúrgicas y 3 de biopsias endoscópicas) en placas recubiertas de colágeno tipo I con medio suplementado para CEB. Al iniciarse la proliferación celular a su alrededor, los explantes se extrajeron y subcultivaron sucesivamente. Las células restantes se dejaron proliferar y se tripsinizaron tras alcanzar más del 50% de confluencia. Se valoraron el número de células obtenidas, la viabilidad y la citoqueratina 7.

Resultados

El número total de células obtenidas con este método superó en una media de 3 veces el número de CEB humanas obtenidas en cultivos primarios simples. El número máximo de subcultivos fue de 5, la viabilidad media (± desviación estándar) fue de 91,9 ± 11,7% y el porcentaje de células positivas para la citoqueratina 7 del 30,71 ± 10,68%.

Conclusiones

El método descrito para amplificar cultivos primarios de CEB permite incrementar la producción de células obtenidas.

Palabras clave:

Cultivos primarios

Células epiteliales bronquiales

Amplificación

Explantes

Citoqueratina 7

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REFERENCES

[1]

VH Velden, HF Versnel.