Expansion of human amniotic epithelial cells using condition cell reprogramming technology
<p dir="ltr">Human amniotic epithelial cells (hAECs) are non-immunogenic epithelial cells that can develop into cells of all three germline lineages. However, a refined clinically reliable method is required to optimize the preparation and banking procedures of hAECs for their succes...
محفوظ في:
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| مؤلفون آخرون: | , , , , |
| منشور في: |
2022
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| الموضوعات: | |
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| _version_ | 1864513520255631360 |
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| author | Aisha Naeem (14157078) |
| author2 | Muhammad Umer Choudhry (18180418) Alex Kroemer (18180421) Simone Wahnschafft (17443838) Wanxing Cui (323677) Chris Albanese (233225) |
| author2_role | author author author author author |
| author_facet | Aisha Naeem (14157078) Muhammad Umer Choudhry (18180418) Alex Kroemer (18180421) Simone Wahnschafft (17443838) Wanxing Cui (323677) Chris Albanese (233225) |
| author_role | author |
| dc.creator.none.fl_str_mv | Aisha Naeem (14157078) Muhammad Umer Choudhry (18180418) Alex Kroemer (18180421) Simone Wahnschafft (17443838) Wanxing Cui (323677) Chris Albanese (233225) |
| dc.date.none.fl_str_mv | 2022-12-31T03:00:00Z |
| dc.identifier.none.fl_str_mv | 10.1007/s13577-022-00849-4 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/Expansion_of_human_amniotic_epithelial_cells_using_condition_cell_reprogramming_technology/25434742 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biological sciences Biochemistry and cell biology Biomedical and clinical sciences Medical biotechnology Amniotic stem cells Amniotic epithelial stem cells Cell culture Human serum Stem cell markers |
| dc.title.none.fl_str_mv | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Human amniotic epithelial cells (hAECs) are non-immunogenic epithelial cells that can develop into cells of all three germline lineages. However, a refined clinically reliable method is required to optimize the preparation and banking procedures of hAECs for their successful translation into clinical studies. With the goal of establishing standardized clinically applicable hAECs cultured cells, we described the use of a powerful epithelial cell culture technique, termed Conditionally Reprogrammed Cells (CRC) for ex vivo expansion of hAECs. The well-established CRC culture method uses a Rho kinase inhibitor (Y-27632) and J2 mouse fibroblast feeder cells to drive the indefinite proliferation of all known epithelial cell types. In this study, we used an optimized CRC protocol to successfully culture hAECs in a CRC medium supplemented with xenogen-free human serum. We established that hAECs thrive under the CRC conditions for over 5 passages while still expressing pluripotent stem markers (OCT-4, SOX-2 and NANOG) and non-immunogenic markers (CD80, CD86 and HLA-G) suggesting that even late-passage hAECs retain their privileged phenotype. The hAECs-CRC cells were infected with a puromycin-selectable lentivirus expressing luciferase and GFP (green fluorescent protein) and stably selected with puromycin. The hAECs expressing GFP were injected subcutaneously into the flanks of Athymic and C57BL6 mice to check the tolerability and stability of cells against the immune system. Chemiluminescence imaging confirmed the presence and viability of cells at days 2, 5, and 42 without acute inflammation or any tumor formation. Collectively, these data indicate that the CRC approach offers a novel solution to expanding hAECs in humanized conditions for future clinical uses, while retaining their primary phenotype.</p><h2>Other Information</h2><p dir="ltr">Published in: Human Cell<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1007/s13577-022-00849-4" target="_blank">https://dx.doi.org/10.1007/s13577-022-00849-4</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_8f4abb06dabcf3a271337f7aea0e4501 |
| identifier_str_mv | 10.1007/s13577-022-00849-4 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/25434742 |
| publishDate | 2022 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Expansion of human amniotic epithelial cells using condition cell reprogramming technologyAisha Naeem (14157078)Muhammad Umer Choudhry (18180418)Alex Kroemer (18180421)Simone Wahnschafft (17443838)Wanxing Cui (323677)Chris Albanese (233225)Biological sciencesBiochemistry and cell biologyBiomedical and clinical sciencesMedical biotechnologyAmniotic stem cellsAmniotic epithelial stem cellsCell cultureHuman serumStem cell markers<p dir="ltr">Human amniotic epithelial cells (hAECs) are non-immunogenic epithelial cells that can develop into cells of all three germline lineages. However, a refined clinically reliable method is required to optimize the preparation and banking procedures of hAECs for their successful translation into clinical studies. With the goal of establishing standardized clinically applicable hAECs cultured cells, we described the use of a powerful epithelial cell culture technique, termed Conditionally Reprogrammed Cells (CRC) for ex vivo expansion of hAECs. The well-established CRC culture method uses a Rho kinase inhibitor (Y-27632) and J2 mouse fibroblast feeder cells to drive the indefinite proliferation of all known epithelial cell types. In this study, we used an optimized CRC protocol to successfully culture hAECs in a CRC medium supplemented with xenogen-free human serum. We established that hAECs thrive under the CRC conditions for over 5 passages while still expressing pluripotent stem markers (OCT-4, SOX-2 and NANOG) and non-immunogenic markers (CD80, CD86 and HLA-G) suggesting that even late-passage hAECs retain their privileged phenotype. The hAECs-CRC cells were infected with a puromycin-selectable lentivirus expressing luciferase and GFP (green fluorescent protein) and stably selected with puromycin. The hAECs expressing GFP were injected subcutaneously into the flanks of Athymic and C57BL6 mice to check the tolerability and stability of cells against the immune system. Chemiluminescence imaging confirmed the presence and viability of cells at days 2, 5, and 42 without acute inflammation or any tumor formation. Collectively, these data indicate that the CRC approach offers a novel solution to expanding hAECs in humanized conditions for future clinical uses, while retaining their primary phenotype.</p><h2>Other Information</h2><p dir="ltr">Published in: Human Cell<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1007/s13577-022-00849-4" target="_blank">https://dx.doi.org/10.1007/s13577-022-00849-4</a></p>2022-12-31T03:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1007/s13577-022-00849-4https://figshare.com/articles/journal_contribution/Expansion_of_human_amniotic_epithelial_cells_using_condition_cell_reprogramming_technology/25434742CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/254347422022-12-31T03:00:00Z |
| spellingShingle | Expansion of human amniotic epithelial cells using condition cell reprogramming technology Aisha Naeem (14157078) Biological sciences Biochemistry and cell biology Biomedical and clinical sciences Medical biotechnology Amniotic stem cells Amniotic epithelial stem cells Cell culture Human serum Stem cell markers |
| status_str | publishedVersion |
| title | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| title_full | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| title_fullStr | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| title_full_unstemmed | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| title_short | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| title_sort | Expansion of human amniotic epithelial cells using condition cell reprogramming technology |
| topic | Biological sciences Biochemistry and cell biology Biomedical and clinical sciences Medical biotechnology Amniotic stem cells Amniotic epithelial stem cells Cell culture Human serum Stem cell markers |