Practical and safe method of cryopreservation for clinical application of human adipose-derived mesenchymal stem cells without a programmable freezer or serum

Gao, Siqiang; Takami, Akiyoshi; Takeshita, Kyosuke; Niwa, Reiko; Kato, Hidefumi; Nakayama, Takayuki

doi:10.1101/664524

Cited by 4 publications

(3 citation statements)

References 19 publications

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“…This could be because of dimethyl sulfoxide (DMSO), which has been used as a cryoprotectant agent at a concentration of 10% for many years. DMSO is able to penetrate cells and inhibit the formation of intracellular ice crystals and membrane rupture (Gao et al, 2020). When compared to DMSO, glycerol has lower viability for the cryopreservation adipose-derived from MSCs.…”

Section: Discussionmentioning

confidence: 99%

Conditioned media and DMSO enhance the cryopreservation of bovine adipose tissue-derived mesenchymal stem cells

Irfan,

Suyatno,

Zulfiqar

et al. 2024

J. Indonesian Trop. Anim. Agric.

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Mesenchymal stem cells derived from adipose tissue (AD-MSCs) show great potential for repro ductive biotechnology in the livestock sector. However, enzyme-based isolation of MSCs is expensive and time-consuming, so it is still rarely done, especially for applications in the livestock sector in de veloping countries. So, MSCs must be cryopreserved with an efficient cryoprotective agent to be stored and reproduced in various laboratories after isolation. This study was aimed to optimize the cryopreser vation media for adipose-derived MSCs in cattle. This study evaluated the viability, proliferation, and morphology of AD-MSCs. The results of this study indicate that a combination of 10% DMSO, 45% DMEM, and 45% conditioned media significantly improves post-thaw viability, proliferation, and sur vival as compared to other mediums. Furthermore, AD-MSCs cryopreserved in this medium exhibit similar morphology as fresh cells. These findings suggest that the optimized cryopreservation medium can enhance the quality and safety of AD-MSCs for clinical applications in the livestock industry.

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Section: Discussionmentioning

confidence: 99%

Conditioned media and DMSO enhance the cryopreservation of bovine adipose tissue-derived mesenchymal stem cells

Irfan,

Suyatno,

Zulfiqar

et al. 2024

J. Indonesian Trop. Anim. Agric.

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“…However, in the case of MSCs and tissue samples, the duration of cryopreservation can be up to 10 years, but this temperature regime does not allow for such a long period of sample preservation [ 10 , 73 ]. A number of studies have demonstrated that with a short storage period (e.g., several months) at −80 °C or at −196 °C, the viability of the thawed MSCs is comparable [ 10 , 74 , 75 ]. In the case of longer cryopreservation (e.g., more than 1 year), it is possible to use freezers that can maintain a stable temperature of about −150 °C, which is an acceptable alternative to liquid nitrogen [ 69 ].…”

Section: Cryopreservation Of Msc Suspensionsmentioning

confidence: 99%

Cryostorage of Mesenchymal Stem Cells and Biomedical Cell-Based Products

Linkova

Rubtsova

Egorikhina

2022

Cells

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Mesenchymal stem cells (MSCs) manifest vast opportunities for clinical use due both to their ability for self-renewal and for effecting paracrine therapeutic benefits. At the same time, difficulties with non-recurrent generation of large numbers of cells due to the necessity for long-term MSC expansion ex vivo, or the requirement for repeated sampling of biological material from a patient significantly limits the current use of MSCs in clinical practice. One solution to these problems entails the creation of a biobank using cell cryopreservation technology. This review is aimed at analyzing and classifying literature data related to the development of protocols for the cryopreservation of various types of MSCs and tissue-engineered structures. The materials in the review show that the existing techniques and protocols for MSC cryopreservation are very diverse, which significantly complicates standardization of the entire process. Here, the selection of cryoprotectors and of cryoprotective media shows the greatest variability. Currently, it is the cryopreservation of cell suspensions that has been studied most extensively, whereas there are very few studies in the literature on the freezing of intact tissues or of tissue-engineered structures. However, even now it is possible to develop general recommendations to optimize the cryopreservation process, making it less traumatic for cells.

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“…Some commercially available cryobags are multiple-chamber CryoStore TM freezing bags (Origen), CryoMACS ® Freezing Bags (Miltenyi Biotec), and Freeze-Pak TM Bio-Containers (CharterMedical) with different fill volumes. Gao et al (2019) reported that human AD-MSCs cryopreserved at -150°C for 24 months in cryobags post-thaw had a viability of >90% with minimal cell clumps with functionality profiles similar to fresh cells. In the case of cryovials, it has been reported that the seal integrity could be compromised for glass vials with rubber stoppers at cryogenic temperatures thus presenting problems of losing product integrity during the critical cryopreservation stage ( Zuleger et al, 2012 ; Hunt, 2019 ).…”

Section: Gmp Based Automated Closed System Manufacturingmentioning

confidence: 99%

Acceleration of Translational Mesenchymal Stromal Cell Therapy Through Consistent Quality GMP Manufacturing

Jayaraman¹,

Lim²,

Ng³

et al. 2021

Front. Cell Dev. Biol.

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Human mesenchymal stromal cell (hMSC) therapy has been gaining immense interest in regenerative medicine and quite recently for its immunomodulatory properties in COVID-19 treatment. Currently, the use of hMSCs for various diseases is being investigated in >900 clinical trials. Despite the huge effort, setting up consistent and robust scalable manufacturing to meet regulatory compliance across various global regions remains a nagging challenge. This is in part due to a lack of definitive consensus for quality control checkpoint assays starting from cell isolation to expansion and final release criterion of clinical grade hMSCs. In this review, we highlight the bottlenecks associated with hMSC-based therapies and propose solutions for consistent GMP manufacturing of hMSCs starting from raw materials selection, closed and modular systems of manufacturing, characterization, functional testing, quality control, and safety testing for release criteria. We also discuss the standard regulatory compliances adopted by current clinical trials to broaden our view on the expectations across different jurisdictions worldwide.

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Practical and safe method of cryopreservation for clinical application of human adipose-derived mesenchymal stem cells without a programmable freezer or serum

Cited by 4 publications

References 19 publications

Conditioned media and DMSO enhance the cryopreservation of bovine adipose tissue-derived mesenchymal stem cells

Conditioned media and DMSO enhance the cryopreservation of bovine adipose tissue-derived mesenchymal stem cells

Cryostorage of Mesenchymal Stem Cells and Biomedical Cell-Based Products

Acceleration of Translational Mesenchymal Stromal Cell Therapy Through Consistent Quality GMP Manufacturing

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