Abstract:
:Pluripotent stem cells offer unique opportunities for curing debilitating diseases. However, further comprehensive research is needed to better understand cell signaling during the differentiation of pluripotent cells into different cell lineages and accordingly to develop clinically applicable protocols. One of the limiting steps for differentiation studies is proper culture and expansion of pluripotent stem cells, which is labor intensive, expensive, and requires a great deal of expertise. This limiting step can be overcome by successful banking and distribution of embryoid bodies (EBs), which are aggregates of pluripotent stem cells and typically the starting point of differentiation protocols. The objective of this study was to investigate the feasibility of EB banking by studying survival and functionality of cryopreserved EBs. To this end, EBs were formed by culturing mouse 129 embryonic stem (ES) cells in the absence of leukemia inhibitory factor (LIF) in hanging drops and then subjected to different cryopreservation protocols. In a series of experiments, we first tested the postthaw survival of EBs as a function of dimethylsulfoxide (DMSO) and extracellular trehalose concentrations and cooling rates. Next, we studied the functionality of cryopreserved EBs by assessing their postthaw attachment, growth, and differentiation into various cell types. Higher (≥5%) DMSO concentrations alone or in combination with trehalose (0.1 M and 0.2 M) yielded good postthaw survival rates of >80%, whereas cooling of EBs at 1°C/min in the presence of 5% DMSO +0.1 M trehalose gave the best attachment and growth rates, with differentiation into cell lineages of three germ layers. Taken together, our results suggest that EBs are tolerant to cryopreservation-associated stresses and retain their differentiation potential after freezing and thawing. Furthermore, our experiments with dissociated EB cells and nondissociated EBs suggest that the extracellular matrix may play a beneficial role in the cryotolerance of EBs. Overall, our data support the feasibility of EB banking, which would facilitate advancement of cell-based therapies.
journal_name
Rejuvenation Resjournal_title
Rejuvenation researchauthors
Sharma S,Szurek EA,Rzucidlo JS,Liour SS,Eroglu Adoi
10.1089/rej.2011.1186subject
Has Abstractpub_date
2011-12-01 00:00:00pages
641-9issue
6eissn
1549-1684issn
1557-8577journal_volume
14pub_type
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journal_title:Rejuvenation research
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journal_title:Rejuvenation research
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pub_type: 杂志文章
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journal_title:Rejuvenation research
pub_type: 杂志文章
doi:10.1089/rej.2008.0666
更新日期:2008-04-01 00:00:00
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journal_title:Rejuvenation research
pub_type: 杂志文章
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journal_title:Rejuvenation research
pub_type: 杂志文章
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journal_title:Rejuvenation research
pub_type: 杂志文章
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journal_title:Rejuvenation research
pub_type: 杂志文章
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journal_title:Rejuvenation research
pub_type: 杂志文章
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journal_title:Rejuvenation research
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journal_title:Rejuvenation research
pub_type: 杂志文章,评审
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更新日期:2005-01-01 00:00:00
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doi:10.1089/rej.2010.1031
更新日期:2010-10-01 00:00:00
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journal_title:Rejuvenation research
pub_type: 杂志文章,评审
doi:10.1089/rej.2006.9.77
更新日期:2006-04-01 00:00:00
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journal_title:Rejuvenation research
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doi:10.1089/rej.2016.1885
更新日期:2017-06-01 00:00:00
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pub_type: 杂志文章,评审
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更新日期:2004-10-01 00:00:00
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journal_title:Rejuvenation research
pub_type: 杂志文章
doi:10.1089/rej.2008.0704
更新日期:2008-06-01 00:00:00