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Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth

PURPOSE: Since the oral environment harbors various microorganisms, the removal of contaminants during the primary culture process of stem cells from human exfoliated deciduous teeth (SHEDs) is very important. We investigated optimal methods for primary culture of SHEDs with minimal contamination ra...

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Autores principales: Lee, Du-man, Lee, Sun Haeng, Hong, Tae Hee, Lee, Jae Cheoun, Nam, Hyun, Joo, Kyeung Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10415792/
https://www.ncbi.nlm.nih.gov/pubmed/37576800
http://dx.doi.org/10.1016/j.jobcr.2023.06.005
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author Lee, Du-man
Lee, Sun Haeng
Hong, Tae Hee
Lee, Jae Cheoun
Nam, Hyun
Joo, Kyeung Min
author_facet Lee, Du-man
Lee, Sun Haeng
Hong, Tae Hee
Lee, Jae Cheoun
Nam, Hyun
Joo, Kyeung Min
author_sort Lee, Du-man
collection PubMed
description PURPOSE: Since the oral environment harbors various microorganisms, the removal of contaminants during the primary culture process of stem cells from human exfoliated deciduous teeth (SHEDs) is very important. We investigated optimal methods for primary culture of SHEDs with minimal contamination rates. MATERIALS AND METHODS: Three different storage conditions for deciduous teeth were utilized:1) storing teeth in Hank's Balanced Salt Solution (HBSS) with 3% penicillin and streptomycin (P/S), 2) storing teeth in HBSS with 3% antibiotics and antimycotics (A-A), and 3) storing teeth in HBSS with A-A, and additional washing with 70% ethanol just before primary culture of dental pulp. In addition, the storage time from the extraction of teeth to the primary culture was measured. RESULTS: The contamination rates were about 70% for HBSS with P/S, 40% for HBSS with A-A, and less than 10% for HBSS with A-A and additional washing with 70% ethanol. When the primary culture was conducted within 12 h after teeth extraction, the contamination rate was the lowest in all conditions. Furthermore, when the teeth were delivered in HBSS with A-A and an additional 70% ethanol washing was performed, the contamination rate was 0% until 48 h after teeth extraction. Ethanol washing had little effect on the cellular characteristics and stemness of SHEDs, including their morphology, growth rate, expression of surface markers, and differentiation potential. CONCLUSIONS: We suggested that both delivering teeth in HBSS with A-A and additional 70% ethanol washing are critical considerations for the successful culture of SHEDs without contamination.
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spelling pubmed-104157922023-08-12 Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth Lee, Du-man Lee, Sun Haeng Hong, Tae Hee Lee, Jae Cheoun Nam, Hyun Joo, Kyeung Min J Oral Biol Craniofac Res Article PURPOSE: Since the oral environment harbors various microorganisms, the removal of contaminants during the primary culture process of stem cells from human exfoliated deciduous teeth (SHEDs) is very important. We investigated optimal methods for primary culture of SHEDs with minimal contamination rates. MATERIALS AND METHODS: Three different storage conditions for deciduous teeth were utilized:1) storing teeth in Hank's Balanced Salt Solution (HBSS) with 3% penicillin and streptomycin (P/S), 2) storing teeth in HBSS with 3% antibiotics and antimycotics (A-A), and 3) storing teeth in HBSS with A-A, and additional washing with 70% ethanol just before primary culture of dental pulp. In addition, the storage time from the extraction of teeth to the primary culture was measured. RESULTS: The contamination rates were about 70% for HBSS with P/S, 40% for HBSS with A-A, and less than 10% for HBSS with A-A and additional washing with 70% ethanol. When the primary culture was conducted within 12 h after teeth extraction, the contamination rate was the lowest in all conditions. Furthermore, when the teeth were delivered in HBSS with A-A and an additional 70% ethanol washing was performed, the contamination rate was 0% until 48 h after teeth extraction. Ethanol washing had little effect on the cellular characteristics and stemness of SHEDs, including their morphology, growth rate, expression of surface markers, and differentiation potential. CONCLUSIONS: We suggested that both delivering teeth in HBSS with A-A and additional 70% ethanol washing are critical considerations for the successful culture of SHEDs without contamination. Elsevier 2023 2023-07-31 /pmc/articles/PMC10415792/ /pubmed/37576800 http://dx.doi.org/10.1016/j.jobcr.2023.06.005 Text en © 2023 Published by Elsevier B.V. on behalf of Craniofacial Research Foundation. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Lee, Du-man
Lee, Sun Haeng
Hong, Tae Hee
Lee, Jae Cheoun
Nam, Hyun
Joo, Kyeung Min
Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title_full Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title_fullStr Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title_full_unstemmed Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title_short Effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
title_sort effects of ethanol washing and storage duration on primary culture of stem cells from human exfoliated deciduous teeth
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10415792/
https://www.ncbi.nlm.nih.gov/pubmed/37576800
http://dx.doi.org/10.1016/j.jobcr.2023.06.005
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