Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction
We aimed to investigate whether molecular clues from the extracellular matrix (ECM) can induce oral epithelial differentiation of pluripotent stem cells. Mouse embryonic stem cells (ESC) of the feeder-independent cell line E14 were used as a model for pluripotent stem cells. They were first grown in...
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/9/7694 |
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| author | Ridhima Das Lisa Harper Kayoko Kitajima Tarig Al-Hadi Osman Mihaela Roxana Cimpan Anne Chr. Johannssen Salwa Suliman Ian C. Mackenzie Daniela-Elena Costea |
| author_facet | Ridhima Das Lisa Harper Kayoko Kitajima Tarig Al-Hadi Osman Mihaela Roxana Cimpan Anne Chr. Johannssen Salwa Suliman Ian C. Mackenzie Daniela-Elena Costea |
| author_sort | Ridhima Das |
| collection | DOAJ |
| description | We aimed to investigate whether molecular clues from the extracellular matrix (ECM) can induce oral epithelial differentiation of pluripotent stem cells. Mouse embryonic stem cells (ESC) of the feeder-independent cell line E14 were used as a model for pluripotent stem cells. They were first grown in 2D on various matrices in media containing vitamin C and without leukemia inhibitory factor (LIF). Matrices investigated were gelatin, laminin, and extracellular matrices (ECM) synthesized by primary normal oral fibroblasts and keratinocytes in culture. Differentiation into epithelial lineages was assessed by light microscopy, immunocytochemistry, and flow cytometry for cytokeratins and stem cell markers. ESC grown in 2D on various matrices were afterwards grown in 3D organotypic cultures with or without oral fibroblasts in the collagen matrix and examined histologically and by immunohistochemistry for epithelial (keratin pairs 1/10 and 4/13 to distinguish epidermal from oral epithelia and keratins 8,18,19 to phenotype simple epithelia) and mesenchymal (vimentin) phenotypes. ECM synthesized by either oral fibroblasts or keratinocytes was able to induce, in 2D cultures, the expression of cytokeratins of the stratified epithelial phenotype. When grown in 3D, all ESC developed into two morphologically distinct cell populations on collagen gels: (i) epithelial-like cells organized in islands with occasional cyst- or duct-like structures and (ii) spindle-shaped cells suggestive of mesenchymal differentiation. The 3D culture on oral fibroblast-populated collagen matrices was necessary for further differentiation into oral epithelia. Only ESC initially grown on 2D keratinocyte or fibroblast-synthesized matrices reached full epithelial maturation. In conclusion, ESC can generate oral epithelia under matrix instruction. |
| first_indexed | 2024-03-11T04:18:31Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T04:18:31Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d4691a7b46724b979b380125f61409d62023-11-17T22:59:18ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01249769410.3390/ijms24097694Embryonic Stem Cells Can Generate Oral Epithelia under Matrix InstructionRidhima Das0Lisa Harper1Kayoko Kitajima2Tarig Al-Hadi Osman3Mihaela Roxana Cimpan4Anne Chr. Johannssen5Salwa Suliman6Ian C. Mackenzie7Daniela-Elena Costea8Gade Laboratory for Pathology and Center for Cancer Biomarkers CCBIO, Institute for Clinical Medicine, University of Bergen, 5020 Bergen, NorwayInstitute for Cell and Molecular Science, Queen Mary University of London, London E1 4NS, UKDepartment of Endodontics, The Nippon Dental University School of Life Dentistry at Niigata, Niigata 951-8580, JapanDepartment of Clinical Dentistry, University of Bergen, 5020 Bergen, NorwayDepartment of Clinical Dentistry, University of Bergen, 5020 Bergen, NorwayGade Laboratory for Pathology and Center for Cancer Biomarkers CCBIO, Institute for Clinical Medicine, University of Bergen, 5020 Bergen, NorwayDepartment of Clinical Dentistry, University of Bergen, 5020 Bergen, NorwayInstitute for Cell and Molecular Science, Queen Mary University of London, London E1 4NS, UKGade Laboratory for Pathology and Center for Cancer Biomarkers CCBIO, Institute for Clinical Medicine, University of Bergen, 5020 Bergen, NorwayWe aimed to investigate whether molecular clues from the extracellular matrix (ECM) can induce oral epithelial differentiation of pluripotent stem cells. Mouse embryonic stem cells (ESC) of the feeder-independent cell line E14 were used as a model for pluripotent stem cells. They were first grown in 2D on various matrices in media containing vitamin C and without leukemia inhibitory factor (LIF). Matrices investigated were gelatin, laminin, and extracellular matrices (ECM) synthesized by primary normal oral fibroblasts and keratinocytes in culture. Differentiation into epithelial lineages was assessed by light microscopy, immunocytochemistry, and flow cytometry for cytokeratins and stem cell markers. ESC grown in 2D on various matrices were afterwards grown in 3D organotypic cultures with or without oral fibroblasts in the collagen matrix and examined histologically and by immunohistochemistry for epithelial (keratin pairs 1/10 and 4/13 to distinguish epidermal from oral epithelia and keratins 8,18,19 to phenotype simple epithelia) and mesenchymal (vimentin) phenotypes. ECM synthesized by either oral fibroblasts or keratinocytes was able to induce, in 2D cultures, the expression of cytokeratins of the stratified epithelial phenotype. When grown in 3D, all ESC developed into two morphologically distinct cell populations on collagen gels: (i) epithelial-like cells organized in islands with occasional cyst- or duct-like structures and (ii) spindle-shaped cells suggestive of mesenchymal differentiation. The 3D culture on oral fibroblast-populated collagen matrices was necessary for further differentiation into oral epithelia. Only ESC initially grown on 2D keratinocyte or fibroblast-synthesized matrices reached full epithelial maturation. In conclusion, ESC can generate oral epithelia under matrix instruction.https://www.mdpi.com/1422-0067/24/9/7694mouseembryonic stem cellsdifferentiationskinoral mucosamatrix |
| spellingShingle | Ridhima Das Lisa Harper Kayoko Kitajima Tarig Al-Hadi Osman Mihaela Roxana Cimpan Anne Chr. Johannssen Salwa Suliman Ian C. Mackenzie Daniela-Elena Costea Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction International Journal of Molecular Sciences mouse embryonic stem cells differentiation skin oral mucosa matrix |
| title | Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction |
| title_full | Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction |
| title_fullStr | Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction |
| title_full_unstemmed | Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction |
| title_short | Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction |
| title_sort | embryonic stem cells can generate oral epithelia under matrix instruction |
| topic | mouse embryonic stem cells differentiation skin oral mucosa matrix |
| url | https://www.mdpi.com/1422-0067/24/9/7694 |
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