Effects of storage media, supplements and cryopreservation methods on quality of stem cells

Erol, Özgür Doğuş; Pervin, Burcu; Şeker, Mehmet Emİn; Aerts‐Kaya, Fatima

doi:10.4252/wjsc.v13.i9.1197

Cited by 21 publications

(16 citation statements)

References 123 publications

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“…Specifically, it was found that an apparent >90% viable population 1 h post-thaw decreased to less than half that (between 20–40%) by 24 h post-thaw following hHPC cryopreservation using traditional culture media with 5 or 10% DMSO ( Figure 1 ). This finding was consistent with previous reports that describe HPC and/or HSC post-thaw viability to range between 40 and 60% of pre-freeze controls with varying cryopreservation media formulations and cryoprotective agents, including 10% DMSO [ 64 , 88 , 95 , 96 , 97 , 98 , 99 , 100 ]. When samples were cryopreserved using Unisol™, overall hHPC viability at 24 h post-thaw increased to 60–65% depending on the DMSO concentration ( Figure 2 ).…”

Section: Discussionsupporting

confidence: 92%

“…To this end, initial observations of the CD34+ cells within the surviving HPC samples suggest a similar % fraction between samples cryopreserved in media or Unisol + DMSO and cultured in recovery media with or without n-acetyl cysteine supplementation. While preliminary, these initial findings are consistent with previous reports [ 51 , 88 , 95 , 96 , 97 , 98 , 99 , 100 , 101 ], and suggest that post-thaw conditioning may provide for increased recovery of total viable cells as well as maintenance of the CD34+ subpopulation, correlating to a higher number of viable CD34+ cells post-thaw. While encouraging, these observations are highly preliminary; therefore, further investigation must be completed prior to drawing any definitive conclusions.…”

Section: Discussionsupporting

confidence: 90%

“…Reports have shown that a 50 to 75% reduction in overall cell viability can result in the initial 24 h post-thaw period in numerous cell systems [ 22 , 94 ]. However, detailed reports on the extent of hHPC cell loss over this interval are limited [ 88 , 95 , 96 , 97 , 98 , 99 , 100 ]. Our data illustrated that there was a substantial level of hHPC loss over the initial 24 h recovery period ( Figure 1 ).…”

Section: Discussionmentioning

confidence: 99%

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Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model

Baust

Snyder

Buskirk

et al. 2022

Cells

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The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, storage, and distribution of living materials. Despite serving a critical role and substantial improvements over the last several decades, cryopreservation often remains a bottleneck impacting numerous areas including cell therapy, tissue engineering, and tissue banking. Studies have illustrated the impact and benefit of controlling cryopreservation-induced delayed-onset cell death (CIDOCD) through various “front end” strategies, such as specialized media, new cryoprotective agents, and molecular control during cryopreservation. While proving highly successful, a substantial level of cell death and loss of cell function remains associated with cryopreservation. Recently, we focused on developing technologies (RevitalICE™) designed to reduce the impact of CIDOCD through buffering the cell stress response during the post-thaw recovery phase in an effort to improve the recovery of previously cryopreserved samples. In this study, we investigated the impact of modulating apoptotic caspase activation, oxidative stress, unfolded protein response, and free radical damage in the initial 24 h post-thaw on overall cell survival. Human hematopoietic progenitor cells in vitro cryopreserved in both traditional extracellular-type and intracellular-type cryopreservation freeze media were utilized as a model cell system to assess impact on survival. Our findings demonstrated that through the modulation of several of these pathways, improvements in cell recovery were obtained, regardless of the freeze media and dimethyl sulfoxide concentration utilized. Specifically, through the use of oxidative stress inhibitors, an average increase of 20% in overall viability was observed. Furthermore, the results demonstrated that by using the post-thaw recovery reagent on samples cryopreserved in intracellular-type media (Unisol™), improvements in overall cell survival approaching 80% of non-frozen controls were attained. While improvements in overall survival were obtained, an assessment on the impact of specific cell subpopulations and functionality remains to be completed. While work remains, these results represent an important step forward in the development of improved cryopreservation processes for use in discovery science, and commercial and clinical settings.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model

Baust

Snyder

Buskirk

et al. 2022

Cells

View full text Add to dashboard Cite

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“…The toxicity of most common CPAs limits their application in medical protocols of cryopreservation, including those intended for cell therapy and tissue/organ transplantation of immune hematology, regenerative medicine, and the reproductive biology area of competence. These aspects have extensively been described in other recent review articles (see for example [ 3 , 25 , 26 , 27 ]) and will find limited space in this mini-review article; for this reason, we apologize to all the authors that have not been cited in our article even if they have given important scientific contributions to this field.…”

Section: Introductionmentioning

confidence: 98%

Natural Cryoprotective and Cytoprotective Agents in Cryopreservation: A Focus on Melatonin

et al. 2022

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Cryoprotective and cytoprotective agents (Cytoprotective Agents) are fundamental components of the cryopreservation process. This review presents the essentials of the cryopreservation process by examining its drawbacks and the role of cytoprotective agents in protecting cell physiology. Natural cryoprotective and cytoprotective agents, such as antifreeze proteins, sugars and natural deep eutectic systems, have been compared with synthetic ones, addressing their mechanisms of action and efficacy of protection. The final part of this article focuses melatonin, a hormonal substance with antioxidant properties, and its emerging role as a cytoprotective agent for somatic cells and gametes, including ovarian tissue, spermatozoa and spermatogonial stem cells.

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“…The total substitute of DMSO is not advisable, as it affects the viability and decreases the shelf life of MSCs during cryopreservation. Though many in-house cryopreservation formulations have been reported [19,20], it is important that all their constituents meet the bio-safety standards. The use of approved and commercially available freezing formulations will ease the complications and facilitates hassle-free filing to NDA (New Drug Application), auditing, screening and testing etc.…”

Section: Discussionmentioning

confidence: 99%

Evaluation and Characterization of Human Bone Marrow Mesenchymal Stromal Cells Cryopreserved in Animal Component-Free, Chemically Defined, Serum-Free Conditions

Kannan¹,

Bhagwat²,

Gupta³

et al. 2023

Possibilities and Limitations in Current Translational Stem Cell Research

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Mesenchymal stromal cells (MSCs) have the potential to treat various disease indications and are the future of cell therapy-based regenerative medicine. Typically, MSCs cryopreserved in serum-containing freezing formulation are supplied at the clinical site, which necessities that this formulation is removed before the administration. This is a cumbersome process, and there is an immediate need for identifying serum-free, xeno-free cryopreservation medium that can be readily used. Here, we analysed two commercially available serum-free, xeno-free, defined freezing media viz., CryoStor 5 (CS5) and CryoStor 10 (CS10) on their effect on human bone marrow MSCs at different freezing cell densities (5, 10, 12.5, 15 and 25 million cells per ml) over a period of 6 months and compared them to the in-house PlasmaLyte A (PLA)-based cryopreservation media. We found that the MSCs cryopreserved in CS5 and CS10 showed similar characteristics as compared with the in-house freezing media for the various parameters analysed including post-thaw recovery, viability, phenotypic marker expression, CFU-F ability and trilineage differentiation potential of the MSCs. Our results show that human MSC could be successfully cryopreserved using serum-free and xeno-free cryopreservation media and can be delivered to the bedside without any manipulations.

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Effects of storage media, supplements and cryopreservation methods on quality of stem cells

Cited by 21 publications

References 123 publications

Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model

Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model

Natural Cryoprotective and Cytoprotective Agents in Cryopreservation: A Focus on Melatonin

Evaluation and Characterization of Human Bone Marrow Mesenchymal Stromal Cells Cryopreserved in Animal Component-Free, Chemically Defined, Serum-Free Conditions

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