Comparison of different cryopreservation protocols for human umbilical cord tissue as source of mesenchymal stem cells

Işildar, Başak; Özkan, Serbay; Öncül, Mahmut; Başlar, Zafer; Kaleli, Semih; Taşyürekli, Mustafa; Koyutürk, Meral

doi:10.1016/j.acthis.2019.02.008

Cited by 19 publications

(19 citation statements)

References 35 publications

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“…PDT analysisThe defined doses of H 2 O 2 and RES were applied to 24-hour cultured cells for 48 hours. After the application, the cells were collected and counted in the Neubauer-Improved (Marienfield) hemocytometer.PDT was calculated by following formulas18 : Population doubling = log total cell number=cultured cell number ð Þ × 3:33 Population doubling time = Total time=population doubling 2.4 | Immunocytochemistry staining At the last hour of experiments (H 2 O 2 and RES exposure) BrdU (20 mM) was added to the medium and incubated for 60 minutes at 37 C. In the end of 48th hour based on previously established protocols, 20 the cells were fixed and routine immunocytochemical procedures were applied.…”

mentioning

confidence: 99%

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration

Kaleci

Koyutürk

2020

Dermatologic Therapy

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confidence: 99%

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration

Kaleci

Koyutürk

2020

Dermatologic Therapy

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“…The long-term preservation of ADSCs is crucial for clinical applications of cell-based therapy. Cryopreservation is commonly used to preserve cells, but cryodamage during the freezing process is a threat to cell viability [ 23 ]. CPAs help minimize the formation of ice crystals and other cryodamage by reducing the crystallization process of water and increasing the viscosity of the solution [ 13 , 24 , 25 ].…”

Section: Discussionmentioning

confidence: 99%

The combination of trehalose and glycerol: an effective and non-toxic recipe for cryopreservation of human adipose-derived stem cells

et al. 2020

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Background Adipose-derived stem cells (ADSCs) promote tissue regeneration and repair. Cryoprotective agents (CPAs) protect cells from cryodamage during cryopreservation. Safe and efficient cryopreservation of ADSCs is critical for cell-based therapy in clinical applications. However, most CPAs are used at toxic concentrations, limiting their clinical application. Objective The aim of this study is to develop a non-toxic xeno-free novel CPA aiming at achieving high-efficiency and low-risk ADSC cryopreservation. Methods We explored different concentrations of trehalose (0.3 M, 0.6 M, 1.0 M, and 1.25 M) and glycerol (10%, 20%, and 30% v/v) for optimization and evaluated and compared the outcomes of ADSCs cryopreservation between a combination of trehalose and glycerol and the commonly used CPA DMSO (10%) + FBS (90%). All samples were slowly frozen and stored in liquid nitrogen for 30 days. The effectiveness was evaluated by the viability, proliferation, migration, and multi-potential differentiation of the ADSCs after thawing. Results Compared with the groups treated with individual reagents, the 1.0 M trehalose (Tre) + 20% glycerol (Gly) group showed significantly higher efficiency in preserving ADSC activities after thawing, with better outcomes in both cell viability and proliferation capacity. Compared with the 10% DMSO + 90% FBS treatment, the ADSCs preserved in 1.0 M Tre + 20% Gly showed similar cell viability, surface markers, and multi-potential differentiation but a significantly higher migration capability. The results indicated that cell function preservation can be improved by 1.0 M Tre + 20% Gly. Conclusions The 1.0 M Tre + 20% Gly treatment preserved ADSCs with a higher migration capability than 10% DMSO + 90% FBS and with viability higher than that with trehalose or glycerol alone but similar to that with 10% DMSO + 90% FBS and fresh cells. Moreover, the new CPA achieves stemness and multi-potential differentiation similar to those in fresh cells. Our results demonstrate that 1.0 M Tre + 20% Gly can more efficiently cryopreserve ADSCs and is a non-toxic CPA that may be suitable for clinical applications.

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“…The long-term preservation of ADSCs is crucial for clinical applications of cell-based therapy. Cryopreservation is commonly used to preserve cells, but cryodamage during the freezing process is a threat to cell viability [23]. CPAs help minimize the formation of ice crystals and other cryodamage by reducing the crystallization process of water and increasing the viscosity of the solution [24][25][26].…”

Section: Discussionmentioning

confidence: 99%

The Combination of Trehalose and Glycerol: An Effective and Non-Toxic Recipe for Cryopreservation of Human Adipose-Derived Stem Cells

Zhang¹,

Tan²,

Xie³

et al. 2020

Preprint

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Background: Adipose-derived stem cells (ADSCs) promote tissue regeneration and repair. Cryoprotective agents (CPAs) protect cells from cryodamage during cryopreservation. Safe and efficient cryopreservation of ADSCs is critical for cell-based therapy in clinical applications. However, most CPAs are used at toxic concentrations, limiting their clinical application. Objective: The aim of this study is to develop a non-toxic xeno-free novel CPA aiming at achieving high-efficiency and low-risk ADSC cryopreservation.Methods: We explored different concentrations of trehalose (0.3 M, 0.6 M, 1.0 M, and 1.25 M) and glycerol (10%, 20%, and 30% v/v) for optimization and evaluated and compared the outcomes of ADSCs cryopreservation between a combination of trehalose and glycerol and the commonly used CPA DMSO (10%) + FBS (90%). All samples were slowly frozen and stored in liquid nitrogen for 30 days. The effectiveness was evaluated by the viability, proliferation, migration and multi-potential differentiation of the ADSCs after thawing. Results: Compared with the groups treated with individual reagents, the 1.0 M Tre + 20% Gly group showed significantly higher efficiency in preserving ADSC activities after thawing, with better outcomes in both cell viability and proliferation capacity. Compared with the 10% DMSO + 90% FBS treatment, the ADSCs preserved in 1.0 M Tre + 20% Gly showed similar cell viability, surface markers and multi-potential differentiation but a significantly higher migration capability. The results indicated that cell function preservation can be improved by 1.0 M Tre + 20% Gly. Conclusions: The 1.0 M Tre + 20% Gly treatment preserved ADSCs with a higher migration capability than 10% DMSO + 90% FBS and with viability higher than that with trehalose or glycerol alone but similar to that with 10% DMSO + 90% FBS and fresh cells. Moreover, the new CPA achieves stemness and multi-potential differentiation similar to those in fresh cells. Our results demonstrate that 1.0 M Tre + 20% Gly can more efficiently cryopreserve ADSCs and is a non-toxic CPA that may be suitable for clinical applications.

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Comparison of different cryopreservation protocols for human umbilical cord tissue as source of mesenchymal stem cells

Cited by 19 publications

References 35 publications

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration

Efficacy of resveratrol in the wound healing process by reducing oxidative stress and promoting fibroblast cell proliferation and migration

The combination of trehalose and glycerol: an effective and non-toxic recipe for cryopreservation of human adipose-derived stem cells

The Combination of Trehalose and Glycerol: An Effective and Non-Toxic Recipe for Cryopreservation of Human Adipose-Derived Stem Cells

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