A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion
We aimed to investigate whether a novel technique of human amniotic membrane (HAM) preparation that mimics the crypts in the limbus enhances the number of progenitor cells cultured ex vivo. The HAMs were sutured on polyester membrane (1) standardly, to obtain a flat HAM surface, or (2) loosely, achi...
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| Format: | Article |
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MDPI AG
2023-02-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/5/738 |
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| author | Jovana Bisevac Morten Carstens Moe Liv Drolsum Olav Kristianslund Goran Petrovski Agate Noer |
| author_facet | Jovana Bisevac Morten Carstens Moe Liv Drolsum Olav Kristianslund Goran Petrovski Agate Noer |
| author_sort | Jovana Bisevac |
| collection | DOAJ |
| description | We aimed to investigate whether a novel technique of human amniotic membrane (HAM) preparation that mimics the crypts in the limbus enhances the number of progenitor cells cultured ex vivo. The HAMs were sutured on polyester membrane (1) standardly, to obtain a flat HAM surface, or (2) loosely, achieving the radial folding to mimic crypts in the limbus. Immunohistochemistry was used to demonstrate a higher number of cells positive for progenitor markers p63α (37.56 ± 3.34% vs. 62.53 ± 3.32%, <i>p</i> = 0.01) and SOX9 (35.53 ± 0.96% vs. 43.23 ± 2.32%, <i>p</i> = 0.04), proliferation marker Ki-67 (8.43 ± 0.38 % vs. 22.38 ± 1.95 %, <i>p</i> = 0.002) in the crypt-like HAMs vs. flat HAMs, while no difference was found for the quiescence marker CEBPD (22.99 ± 2.96% vs. 30.49 ± 3.33 %, <i>p</i> = 0.17). Most of the cells stained negative for the corneal epithelial differentiation marker KRT3/12, and some were positive for N-cadherin in the crypt-like structures, but there was no difference in staining for E-cadherin and CX43 in crypt-like HAMs vs. flat HAMs. This novel HAM preparation method enhanced the number of progenitor cells expanded in the crypt-like HAM compared to cultures on the conventional flat HAM. |
| first_indexed | 2024-03-11T07:28:46Z |
| format | Article |
| id | doaj.art-827d098c92384c3f9b931db8a8d97099 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T07:28:46Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-827d098c92384c3f9b931db8a8d970992023-11-17T07:27:29ZengMDPI AGCells2073-44092023-02-0112573810.3390/cells12050738A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon ExpansionJovana Bisevac0Morten Carstens Moe1Liv Drolsum2Olav Kristianslund3Goran Petrovski4Agate Noer5Center for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Oslo University Hospital, P.O. Box 4956 Nydalen, 0424 Oslo, NorwayCenter for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Oslo University Hospital, P.O. Box 4956 Nydalen, 0424 Oslo, NorwayInstitute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1171 Blindern, 0318 Oslo, NorwayInstitute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1171 Blindern, 0318 Oslo, NorwayCenter for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Oslo University Hospital, P.O. Box 4956 Nydalen, 0424 Oslo, NorwayCenter for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Oslo University Hospital, P.O. Box 4956 Nydalen, 0424 Oslo, NorwayWe aimed to investigate whether a novel technique of human amniotic membrane (HAM) preparation that mimics the crypts in the limbus enhances the number of progenitor cells cultured ex vivo. The HAMs were sutured on polyester membrane (1) standardly, to obtain a flat HAM surface, or (2) loosely, achieving the radial folding to mimic crypts in the limbus. Immunohistochemistry was used to demonstrate a higher number of cells positive for progenitor markers p63α (37.56 ± 3.34% vs. 62.53 ± 3.32%, <i>p</i> = 0.01) and SOX9 (35.53 ± 0.96% vs. 43.23 ± 2.32%, <i>p</i> = 0.04), proliferation marker Ki-67 (8.43 ± 0.38 % vs. 22.38 ± 1.95 %, <i>p</i> = 0.002) in the crypt-like HAMs vs. flat HAMs, while no difference was found for the quiescence marker CEBPD (22.99 ± 2.96% vs. 30.49 ± 3.33 %, <i>p</i> = 0.17). Most of the cells stained negative for the corneal epithelial differentiation marker KRT3/12, and some were positive for N-cadherin in the crypt-like structures, but there was no difference in staining for E-cadherin and CX43 in crypt-like HAMs vs. flat HAMs. This novel HAM preparation method enhanced the number of progenitor cells expanded in the crypt-like HAM compared to cultures on the conventional flat HAM.https://www.mdpi.com/2073-4409/12/5/738limbusHAMcrypt (CLETLESCprogenitordifferentiation |
| spellingShingle | Jovana Bisevac Morten Carstens Moe Liv Drolsum Olav Kristianslund Goran Petrovski Agate Noer A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion Cells limbus HAM crypt (CLET LESC progenitor differentiation |
| title | A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion |
| title_full | A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion |
| title_fullStr | A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion |
| title_full_unstemmed | A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion |
| title_short | A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion |
| title_sort | novel technique of amniotic membrane preparation mimicking limbal epithelial crypts enhances the number of progenitor cells upon expansion |
| topic | limbus HAM crypt (CLET LESC progenitor differentiation |
| url | https://www.mdpi.com/2073-4409/12/5/738 |
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