The effects of freeze/thawing process on cryopreserved equine umbilical cord blood-derived mesenchymal stem cells

Eini, Fatemeh; Foroutan, Tahereh; Bidadkosh, Arash; Barin, Abbas; Dehghan, Mohammad Mehdi; Tajik, Parviz

doi:10.1007/s00580-011-1355-8

Cited by 8 publications

(9 citation statements)

References 20 publications

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“…In the current study, cell viability was ∼70% after thawing and harvest compared with ∼100% viability of the freshly harvested, continuously cultured cells. These are comparable to experiences in other studies, as are the freezing medium and the freeze-thaw technique used [11][12][13][14]. As such, there is no obvious discrepancy in the technical approaches.…”

Section: Discussionsupporting

confidence: 85%

“…The frozen MSCs, suspended at the desired concentration in a cryopreservation medium suitable for infusion, are then thawed just prior to administration. However, a significant number of the frozen MSCs may undergo apoptosis during the freeze-thaw process, although much subsequent study has attempted to minimize this occurrence with improved freeze-thaw approaches and cryopreservatives [11][12][13][14]. A recent report demonstrated that freshly thawed MSCs were not as effective in in vitro potency assays as continuously cultured MSCs of the same passage number [15].…”

Section: Introductionmentioning

confidence: 99%

“…The length of freezing prior to thaw in the current studies was relatively short (7 days) and may not have had the same effect as longer freezing. Longer freeze time prior to thawing and use has been demonstrated to have detrimental effects on initial cell viability after thawing and on proliferative and differentiation capacities [11][12][13][14]. However, length of freeze time on anti-inflammatory actions is not yet well understood.…”

mentioning

confidence: 99%

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Freshly Thawed and Continuously Cultured Human Bone Marrow-Derived Mesenchymal Stromal Cells Comparably Ameliorate Allergic Airways Inflammation in Immunocompetent Mice

Cruz

Borg

Goodwin

et al. 2015

Stem Cells Translational Medicine

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Recent data suggest that freshly thawed previously frozen mesenchymal stromal cells (MSCs) may not have the same effectiveness or breadth of anti-inflammatory activities as do continuously cultured MSCs. This has significant implications for clinical use, in which many infusion schemes use frozen cells thawed at the bedside for administration. The available data, however, predominantly evaluate in vitro MSC properties, and so far there has been limited in vivo analysis. To further assess this issue, we compared freshly thawed (thawed) versus continuously cultured (fresh) human bone marrowderived MSC (hMSC) administration in a mouse model of mixed Th2/Th17 allergic airway inflammation induced by Aspergillus hyphal extract (AHE) exposures in immunocompetent C57Bl/6 mice. Control cell populations included fresh versus thawed murine bone marrow-derived MSCs (mMSCs) and human lung fibroblasts (HLFs). Systemic administration of both thawed and fresh hMSCs and mMSCs, but not HLFs, at the onset of antigen challenge in previously sensitized mice significantly ameliorated the AHE-provoked increases in airway hyper-reactivity, lung inflammation, and antigenspecific CD4 T-cell Th2 and Th17 phenotype. Notably, there was no difference in effects of fresh versus thawed hMSCs or mMSCs on any outcome measured except for some variability in the effects on the bronchoalveolar lavage fluid composition. These results demonstrated potent xenogeneic effects of human MSCs in an immunocompetent mouse model of allergic airways inflammation and that thawed MSCs are as effective as fresh MSCs. The question of fresh versus thawed MSC effectiveness needs to be investigated carefully and may differ in different in vivo disease-specific models. STEM CELLS TRANSLATIONAL MEDICINE 2015;4:615-624 SIGNIFICANCEThis study addressed whether freshly thawed mesenchymal stromal cells (MSCs) are as effective in in vivo settings as those that have been continuously cultured. It also provided further data demonstrating that xenogeneic use of MSCs in immunocompetent mice is as effective as murine MSCs. This information provides further support and direction for potential clinical use of MSCs in patients with severe asthma.

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Section: Discussionsupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Freshly Thawed and Continuously Cultured Human Bone Marrow-Derived Mesenchymal Stromal Cells Comparably Ameliorate Allergic Airways Inflammation in Immunocompetent Mice

Cruz

Borg

Goodwin

et al. 2015

Stem Cells Translational Medicine

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“…Cell viability decreases from about 80.4% for P1 to 51.2% for P10. 24 A study to assess cooling rate, end temperature, hold time, and thawing rate on human ASC cell membrane integrity showed a significant effect of thawing rate on only P3 and P4 while the interaction between cooling rate and end temperature was significant for P0-P4. 27 Results of studies to investigate effects of cell concentration on post-cryopreservation viability vary.…”

Section: Ell P a S S Age N Umb Er C Once Ntra Ti On A Nd C Ryopmentioning

confidence: 99%

“…There is a direct relationship between equine umbilical cord blood MSC viability and pre‐cryopreservation passage number following cryopreservation in 20% DMEM, 70% FBS, and 10% DMSO for 8 weeks. Cell viability decreases from about 80.4% for P1 to 51.2% for P10 . A study to assess cooling rate, end temperature, hold time, and thawing rate on human ASC cell membrane integrity showed a significant effect of thawing rate on only P3 and P4 while the interaction between cooling rate and end temperature was significant for P0‐P4 …”

Section: Cell Passage Number Concentration and Cryopreservation Durmentioning

confidence: 99%

Adult multipotent stromal cell cryopreservation: Pluses and pitfalls

Duan

Lopez

Hicok³

2017

Veterinary Surgery

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Study and clinical testing of adult multipotent stromal cells (MSCs) are central to progressive improvements in veterinary regenerative medicine. Inherent limitations to long‐term culture preclude use for storage. Until cell line creation from primary isolates becomes routine, MSC stasis at cryogenic temperatures is required for this purpose. Many protocols and reagents, including cryoprotectants, used for veterinary MSCs are derived from those for human and rodent cells. Dissimilarities in cryopreservation strategies play a role in variable MSC behaviors. Familiarity with contemporary cryopreservation reagents and processes is essential to an appreciation of their impact on MSC survival and post‐cryopreservation behavior. In addition to these points, this review includes a brief history and description of current veterinary stem cell regulation.

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The significance of cell-related challenges in the clinical application of tissue engineering

Almela

Brook

Moharamzadeh

2016

J. Biomed. Mater. Res.

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Tissue engineering is increasingly being recognized as a new approach that could alleviate the burden of tissue damage currently managed with transplants or synthetic devices. Making this novel approach available in the future for patients who would potentially benefit is largely dependent on understanding and addressing all those factors that impede the translation of this technology to the clinic. Cell‐associated factors in particular raise many challenges, including those related to cell sources, up‐ and downstream techniques, preservation, and the creation of in vitro microenvironments that enable cells to grow and function as far as possible as they would in vivo. This article highlights the main confounding issues associated with cells in tissue engineering and how these issues may hinder the advancement of therapeutic tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3157–3163, 2016.

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The effects of freeze/thawing process on cryopreserved equine umbilical cord blood-derived mesenchymal stem cells

Cited by 8 publications

References 20 publications

Freshly Thawed and Continuously Cultured Human Bone Marrow-Derived Mesenchymal Stromal Cells Comparably Ameliorate Allergic Airways Inflammation in Immunocompetent Mice

Freshly Thawed and Continuously Cultured Human Bone Marrow-Derived Mesenchymal Stromal Cells Comparably Ameliorate Allergic Airways Inflammation in Immunocompetent Mice

Adult multipotent stromal cell cryopreservation: Pluses and pitfalls

The significance of cell-related challenges in the clinical application of tissue engineering

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