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Evaluation of the quality of non-cultured autologous skin cell suspension after cryopreservation
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Abstract
Autologous non-cultured skin cell therapy has gradually become a potential approach for the treatment of wounds and extensive deep burns [1, 2]. The use of non-cultured skin cell suspensions has been proven to positively influence the wound healing process. Standardizing the cryopreservation procedure for these suspensions holds significant scientific and clinical value.
Objective: To evaluate cell quality after 4 weeks of cryopreservation at different DMSO concentrations using blue Trypan staining and cell culture methods.
Materials and methods: Ten thin split-thickness skin samples (0.25 mm) were collected from donor sites during autologous skin graft surgery. The isolated cell suspensions were cryopreserved at -800C using different DMSO concentrations (9%, 10%, 11%). After thawing, cell viability was assessed using blue Trypan staining, and cultured to evaluate their ability of the cells to attach and proliferate.
Results: The mean post-isolation cell viability was above 80%, with the highest reaching 92.1%. A DMSO concentration of 10% (C2) was identified as optimal for cryopreservation. After 4 weeks of storage, cell viability slightly decreased, but the cells remained viable, adherent, and capable of proliferation in culture.
Conclusion: The study demonstrated that non-cultured autologous skin cell suspensions maintained high viability after 4 weeks of cryopreservation. Cells thawed and cultured in defined media remained viable after 6 days, with 10% DMSO identified as the optimal cryoprotectant concentration.
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Keywords
Skin cell suspension, DMSO, cryopreservation
References
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