Hypothermic Storage of Human Umbilical Vein Endothelial Cells and Their Hydrogel Constructs

Zhang, Xiao-Zhang; Cao, Yuan; Zhao, Gang

doi:10.1089/bio.2019.0105

Cited by 6 publications

(4 citation statements)

References 27 publications

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“…Another report showed poor osteogenic capacity of tissue-engineered bone preserved in an osteogenic medium at 4 • C [45]. Lower survival of MSCs at 4 • C compared to 25 • C has been shown for encapsulated cell aggregates [46]. We showed around 90% recovery of WJ-MSCs after 6 d storage at 25 • C, which we believe to be clinically-acceptable values.…”

Section: Discussionsupporting

confidence: 63%

Hypothermic Storage of 3D Cultured Multipotent Mesenchymal Stromal Cells for Regenerative Medicine Applications

et al. 2022

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The regulatory requirements in cell processing, in the choice of a biomaterial scaffold and in quality control analysis, have to be followed in the clinical application of tissue-engineered grafts. Confirmation of sterility during quality control studies requires prolonged storage of the cell-based construct. After storage, preservation of the functional properties of the cells is an important prerequisite if the cells are to be used for cell-based tissue therapies. The study presented here shows the generation of 3D constructs based on Wharton’s jelly multipotent mesenchymal stromal cells (WJ-MSCs) and the clinically-acceptable HyaloFast® scaffold, and the effect of two- and six-day hypothermic storage of 3D cell-based constructs on the functional properties of populated cells. To study the viability, growth, gene expression, and paracrine secretion of WJ-MSCs within the scaffolds before and after storage, xeno-free culture conditions, metabolic, qPCR, and multiplex assays were applied. The WJ-MSCs adhered and proliferated within the 3D HyaloFast®. Our results show different viability of the cells after the 3D constructs have been stored under mild (25 °C) or strong (4 °C) hypothermia. At 4 °C, the significant decrease of metabolic activity of WJ-MSCs was detected after 2 days of storage, with almost complete cell loss after 6 days. In mild hypothermia (25 °C) the decrease in metabolic activity was less remarkable, confirming the suitability of these conditions for cell preservation in 3D environment. The significant changes were detected in gene expression and in the paracrine secretion profile after 2 and 6 days of storage at 25 °C. The results presented in this study are important for the rapid transfer of tissue engineering approaches into clinical applications.

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Section: Discussionsupporting

confidence: 63%

Hypothermic Storage of 3D Cultured Multipotent Mesenchymal Stromal Cells for Regenerative Medicine Applications

et al. 2022

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“…These effects were, for example, comprehensively summarized by Rubinsky et al: In short, exposure to hypothermia has a beneficial effect-it results in a strongly reduced cell metabolism accompanied by reduced energy consumption and reduced oxygen demand, which results in a relatively robust reaction to shortterm hypothermia exposure-but also a number of harmful effects, the most prominent of which are the cell membrane becoming "leaky" due to a lipid phase transition and a reduced activity of ionic pumps, resulting in an osmotic shock. [45,46] It has been hypothesized previously that this osmotic shock can be alleviated by membrane stabilization and reduced ion diffusion via encapsulation of cells in protective hydrogel microcapsules: adipose tissue-derived MSC (AT-MSC) encapsulated in alginate were able to keep their differentiation potential after 2 weeks at 15°C [47] and human umbilical vein endothelial cells encapsulated in alginate with carboxymethyl chitosan showed an 80% viability after 1 week at 4°C [48] In these studies the cells were encapsulated in crosslinked hydrogels, and previous studies investigating cold-storage of cells concentrated on a timeframe of up to 2 weeks. In a very recent study published only this year, [49] the authors also reported that cell-containing alginate beads can be dissolved and used for bioprinting after storage for 1 week at 15°C with a very high viability of the embedded AT-MSC in printed constructs over 14 days; during the culture time they observed a drastic drop in cell number though.…”

Section: Discussionmentioning

confidence: 99%

Bioinks for Space Missions: The Influence of Long‐Term Storage of Alginate‐Methylcellulose‐Based Bioinks on Printability as well as Cell Viability and Function

Windisch

Reinhardt

Duin

et al. 2023

Adv Healthcare Materials

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Bioprinting is considered a key technology for future space missions and is currently being established on the International Space Station (ISS). With the aim to perform bioink production as a critical and resource‐consuming preparatory step already on Earth and transport a bioink cartridge “ready to use” to the ISS, the storability of bioinks is investigated. Hydrogel blends based on alginate and methylcellulose are laden with either green microalgae of the species Chlorella vulgaris or with different human cell lines including immortilized human mesenchymal stem cells, SaOS‐2 and HepG2, as well as with primary human dental pulp stem cells. The bioinks are filled into printing cartridges and stored at 4°C for up to four weeks. Printability of the bioinks is maintained after storage. Viability and function of the cells embedded in constructs bioprinted from the stored bioinks are investigated during subsequent cultivation: The microalgae survive the storage period very well and show no loss of growth and functionality, however a significant decrease is visible for human cells, varying between the different cell types. The study demonstrates that storage of bioinks is in principle possible and is a promising starting point for future research, making complex printing processes more effective and reproducible.

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“…Some groups have explored hypothermic storage using alginate encapsulation. Human umbilical vein endothelial cells (HUVEC) were shown to maintain around 70% viability after 7 days of encapsulation [ 43 ]. After 3 days, 85% of encapsulated MSC(AT) were recovered and were able to reattach to a culture surface [ 23 ].…”

Section: Discussionmentioning

confidence: 99%

Hypothermic Preservation of Adipose-Derived Mesenchymal Stromal Cells as a Viable Solution for the Storage and Distribution of Cell Therapy Products

et al. 2022

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Cell and gene therapies (CGT) have reached new therapeutic targets but have noticeably high prices. Solutions to reduce production costs might be found in CGT storage and transportation since they typically involve cryopreservation, which is a heavily burdened process. Encapsulation at hypothermic temperatures (e.g., 2–8 °C) could be a feasible alternative. Adipose tissue-derived mesenchymal stromal cells (MSC(AT)) expanded using fetal bovine serum (FBS)- (MSC-FBS) or human platelet lysate (HPL)-supplemented mediums (MSC-HPL) were encapsulated in alginate beads for 30 min, 5 days, and 12 days. After bead release, cell recovery and viability were determined to assess encapsulation performance. MSC identity was verified by flow cytometry, and a set of assays was performed to evaluate functionality. MSC(AT) were able to survive encapsulated for a standard transportation period of 5 days, with recovery values of 56 ± 5% for MSC-FBS and 77 ± 6% for MSC-HPL (which is a negligible drop compared to earlier timepoints). Importantly, MSC function did not suffer from encapsulation, with recovered cells showing robust differentiation potential, expression of immunomodulatory molecules, and hematopoietic support capacity. MSC(AT) encapsulation was proven possible for a remarkable 12 day period. There is currently no solution to completely replace cryopreservation in CGT logistics and supply chain, although encapsulation has shown potential to act as a serious competitor.

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Hypothermic Storage of Human Umbilical Vein Endothelial Cells and Their Hydrogel Constructs

Cited by 6 publications

References 27 publications

Hypothermic Storage of 3D Cultured Multipotent Mesenchymal Stromal Cells for Regenerative Medicine Applications

Hypothermic Storage of 3D Cultured Multipotent Mesenchymal Stromal Cells for Regenerative Medicine Applications

Bioinks for Space Missions: The Influence of Long‐Term Storage of Alginate‐Methylcellulose‐Based Bioinks on Printability as well as Cell Viability and Function

Hypothermic Preservation of Adipose-Derived Mesenchymal Stromal Cells as a Viable Solution for the Storage and Distribution of Cell Therapy Products

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