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Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands

Three-dimensional spheroid culture enhances cell-to-cell interactions among stem cells and promotes the expression of stem cell properties; however, subsequent retrieval and delivery of these cells remain a challenge. We fabricated a thermoresponsive fiber-based microwell scaffold by combining elect...

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Autores principales: Hong, Hye Jin, Cho, Jae-Min, Yoon, Yeo-Jun, Choi, DoJin, Lee, Soohyun, Lee, Hwajung, Ahn, Sujeong, Koh, Won-Gun, Lim, Jae-Yol
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9003645/
https://www.ncbi.nlm.nih.gov/pubmed/35422983
http://dx.doi.org/10.1177/20417314221085645
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author Hong, Hye Jin
Cho, Jae-Min
Yoon, Yeo-Jun
Choi, DoJin
Lee, Soohyun
Lee, Hwajung
Ahn, Sujeong
Koh, Won-Gun
Lim, Jae-Yol
author_facet Hong, Hye Jin
Cho, Jae-Min
Yoon, Yeo-Jun
Choi, DoJin
Lee, Soohyun
Lee, Hwajung
Ahn, Sujeong
Koh, Won-Gun
Lim, Jae-Yol
author_sort Hong, Hye Jin
collection PubMed
description Three-dimensional spheroid culture enhances cell-to-cell interactions among stem cells and promotes the expression of stem cell properties; however, subsequent retrieval and delivery of these cells remain a challenge. We fabricated a thermoresponsive fiber-based microwell scaffold by combining electrospinning and hydrogel micropatterning. The resultant scaffold appeared to facilitate the formation of cellular spheroids of uniform size and enabled the expression of more stem cell-secreting growth factor genes (EGF, IGF-1, FGF1, FGF2, and HGF), pluripotent stem cell-related genes (SOX2 and NANOG), and adult epithelial stem cell-related genes (LGR4, LGR5, and LGR6) than salivary gland stem cells in a monolayer culture (SGSC(monolayer)). The spheroids could be retrieved efficiently by decreasing temperature. SGSC-derived spheroid (SGSC(spheroid)) cells were then implanted into the submandibular glands of mice at 2 weeks after fractionated X-ray irradiation at a dose of 7.5 Gy/day. At 16 weeks post-irradiation, restoration of salivary function was detected only in SGSC(spheroid)-implanted mice. The production of submandibular acini specific mucin increased in SGSC(spheroid)-implanted mice, compared with PBS control. More MIST1(+) mature acinar cells were preserved in the SGSC(spheroid)-implanted group than in the PBS control group. Intriguingly, SGSC(spheroid)-implanted mice exhibited greater amelioration of tissue damage and preservation of KRT7(+) terminally differentiated luminal ductal cells than SGSC(monolayer)-implanted mice. The SGSC(spheroid)-implanted mice also showed less DNA damage and apoptotic cell death than the SGSC(monolayer)-implanted mice at 2 weeks post-implantation. Additionally, a significant increase in Ki67(+)AQP5(+) proliferative acinar cells was noted only in SGSC(spheroid)-implanted mice. Our results suggest that a thermoresponsive fiber-based scaffold could be of use to facilitate the production of function-enhanced SGSC(spheroid) cells and their subsequent retrieval and delivery to damaged salivary glands to alleviate radiation-induced apoptotic cell death and promote salivary gland regeneration.
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spelling pubmed-90036452022-04-13 Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands Hong, Hye Jin Cho, Jae-Min Yoon, Yeo-Jun Choi, DoJin Lee, Soohyun Lee, Hwajung Ahn, Sujeong Koh, Won-Gun Lim, Jae-Yol J Tissue Eng Original Article Three-dimensional spheroid culture enhances cell-to-cell interactions among stem cells and promotes the expression of stem cell properties; however, subsequent retrieval and delivery of these cells remain a challenge. We fabricated a thermoresponsive fiber-based microwell scaffold by combining electrospinning and hydrogel micropatterning. The resultant scaffold appeared to facilitate the formation of cellular spheroids of uniform size and enabled the expression of more stem cell-secreting growth factor genes (EGF, IGF-1, FGF1, FGF2, and HGF), pluripotent stem cell-related genes (SOX2 and NANOG), and adult epithelial stem cell-related genes (LGR4, LGR5, and LGR6) than salivary gland stem cells in a monolayer culture (SGSC(monolayer)). The spheroids could be retrieved efficiently by decreasing temperature. SGSC-derived spheroid (SGSC(spheroid)) cells were then implanted into the submandibular glands of mice at 2 weeks after fractionated X-ray irradiation at a dose of 7.5 Gy/day. At 16 weeks post-irradiation, restoration of salivary function was detected only in SGSC(spheroid)-implanted mice. The production of submandibular acini specific mucin increased in SGSC(spheroid)-implanted mice, compared with PBS control. More MIST1(+) mature acinar cells were preserved in the SGSC(spheroid)-implanted group than in the PBS control group. Intriguingly, SGSC(spheroid)-implanted mice exhibited greater amelioration of tissue damage and preservation of KRT7(+) terminally differentiated luminal ductal cells than SGSC(monolayer)-implanted mice. The SGSC(spheroid)-implanted mice also showed less DNA damage and apoptotic cell death than the SGSC(monolayer)-implanted mice at 2 weeks post-implantation. Additionally, a significant increase in Ki67(+)AQP5(+) proliferative acinar cells was noted only in SGSC(spheroid)-implanted mice. Our results suggest that a thermoresponsive fiber-based scaffold could be of use to facilitate the production of function-enhanced SGSC(spheroid) cells and their subsequent retrieval and delivery to damaged salivary glands to alleviate radiation-induced apoptotic cell death and promote salivary gland regeneration. SAGE Publications 2022-04-07 /pmc/articles/PMC9003645/ /pubmed/35422983 http://dx.doi.org/10.1177/20417314221085645 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Article
Hong, Hye Jin
Cho, Jae-Min
Yoon, Yeo-Jun
Choi, DoJin
Lee, Soohyun
Lee, Hwajung
Ahn, Sujeong
Koh, Won-Gun
Lim, Jae-Yol
Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title_full Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title_fullStr Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title_full_unstemmed Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title_short Thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
title_sort thermoresponsive fiber-based microwells capable of formation and retrieval of salivary gland stem cell spheroids for the regeneration of irradiation-damaged salivary glands
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9003645/
https://www.ncbi.nlm.nih.gov/pubmed/35422983
http://dx.doi.org/10.1177/20417314221085645
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