Generation of lung epithelial-like tissue from human embryonic stem cells
<p>Abstract</p> <p>Background</p> <p>Human embryonic stem cells (hESC) have the capacity to differentiate <it>in vivo </it>and <it>in vitro </it>into cells from all three germ lineages. The aim of the present study was to investigate the effect o...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2009-11-01
|
| Series: | Respiratory Research |
| Online Access: | http://respiratory-research.com/content/10/1/105 |
| _version_ | 1811312135172521984 |
|---|---|
| author | Liebaers Inge De Block Gert Van Haute Lindsey Sermon Karen De Rycke Martine |
| author_facet | Liebaers Inge De Block Gert Van Haute Lindsey Sermon Karen De Rycke Martine |
| author_sort | Liebaers Inge |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Human embryonic stem cells (hESC) have the capacity to differentiate <it>in vivo </it>and <it>in vitro </it>into cells from all three germ lineages. The aim of the present study was to investigate the effect of specific culture conditions on the differentiation of hESC into lung epithelial cells.</p> <p>Methods</p> <p>Undifferentiated hESC, grown on a porous membrane in hESC medium for four days, were switched to a differentiation medium for four days; this was followed by culture in air-liquid interface conditions during another 20 days. Expression of several lung markers was measured by immunohistochemistry and by quantitative real-time RT-PCR at four different time points throughout the differentiation and compared to appropriate controls.</p> <p>Results</p> <p>Expression of <it>CC16 </it>and <it>NKX2.1 </it>showed a 1,000- and 10,000- fold increase at day 10 of differentiation. Other lung markers such as <it>SP-C </it>and <it>Aquaporin 5 </it>had the highest expression after twenty days of culture, as well as two markers for ciliated cells, <it>FOXJ1 </it>and <it>β-tubulin IV</it>. The results from qRT-PCR were confirmed by immunohistochemistry on paraffin-embedded samples. Antibodies against CC16, SP-A and SP-C were chosen as specific markers for Clara Cells and alveolar type II cells. The functionality was tested by measuring the secretion of CC16 in the medium using an enzyme immunoassay.</p> <p>Conclusion</p> <p>These results suggest that by using our novel culture protocol hESC can be differentiated into the major cell types of lung epithelial tissue.</p> |
| first_indexed | 2024-04-13T10:30:54Z |
| format | Article |
| id | doaj.art-b9ae8ae015814d0a82c7b039566609c9 |
| institution | Directory Open Access Journal |
| issn | 1465-9921 |
| language | English |
| last_indexed | 2024-04-13T10:30:54Z |
| publishDate | 2009-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj.art-b9ae8ae015814d0a82c7b039566609c92022-12-22T02:50:11ZengBMCRespiratory Research1465-99212009-11-0110110510.1186/1465-9921-10-105Generation of lung epithelial-like tissue from human embryonic stem cellsLiebaers IngeDe Block GertVan Haute LindseySermon KarenDe Rycke Martine<p>Abstract</p> <p>Background</p> <p>Human embryonic stem cells (hESC) have the capacity to differentiate <it>in vivo </it>and <it>in vitro </it>into cells from all three germ lineages. The aim of the present study was to investigate the effect of specific culture conditions on the differentiation of hESC into lung epithelial cells.</p> <p>Methods</p> <p>Undifferentiated hESC, grown on a porous membrane in hESC medium for four days, were switched to a differentiation medium for four days; this was followed by culture in air-liquid interface conditions during another 20 days. Expression of several lung markers was measured by immunohistochemistry and by quantitative real-time RT-PCR at four different time points throughout the differentiation and compared to appropriate controls.</p> <p>Results</p> <p>Expression of <it>CC16 </it>and <it>NKX2.1 </it>showed a 1,000- and 10,000- fold increase at day 10 of differentiation. Other lung markers such as <it>SP-C </it>and <it>Aquaporin 5 </it>had the highest expression after twenty days of culture, as well as two markers for ciliated cells, <it>FOXJ1 </it>and <it>β-tubulin IV</it>. The results from qRT-PCR were confirmed by immunohistochemistry on paraffin-embedded samples. Antibodies against CC16, SP-A and SP-C were chosen as specific markers for Clara Cells and alveolar type II cells. The functionality was tested by measuring the secretion of CC16 in the medium using an enzyme immunoassay.</p> <p>Conclusion</p> <p>These results suggest that by using our novel culture protocol hESC can be differentiated into the major cell types of lung epithelial tissue.</p>http://respiratory-research.com/content/10/1/105 |
| spellingShingle | Liebaers Inge De Block Gert Van Haute Lindsey Sermon Karen De Rycke Martine Generation of lung epithelial-like tissue from human embryonic stem cells Respiratory Research |
| title | Generation of lung epithelial-like tissue from human embryonic stem cells |
| title_full | Generation of lung epithelial-like tissue from human embryonic stem cells |
| title_fullStr | Generation of lung epithelial-like tissue from human embryonic stem cells |
| title_full_unstemmed | Generation of lung epithelial-like tissue from human embryonic stem cells |
| title_short | Generation of lung epithelial-like tissue from human embryonic stem cells |
| title_sort | generation of lung epithelial like tissue from human embryonic stem cells |
| url | http://respiratory-research.com/content/10/1/105 |
| work_keys_str_mv | AT liebaersinge generationoflungepithelialliketissuefromhumanembryonicstemcells AT deblockgert generationoflungepithelialliketissuefromhumanembryonicstemcells AT vanhautelindsey generationoflungepithelialliketissuefromhumanembryonicstemcells AT sermonkaren generationoflungepithelialliketissuefromhumanembryonicstemcells AT deryckemartine generationoflungepithelialliketissuefromhumanembryonicstemcells |