Short/long-term cryopreservation prior to comet assay of whole-blood leukocytes and in vitro-cultured lung fibroblasts. (2nd September 2021)

• Record Type: Journal Article
• Authors: Volobaev, V.; Bach, S.; Shchetnikova, E.; Vdovina, E.; Rosinskiy, A.; Larionov, A.
• Is Part Of: Toxicology mechanisms and methods. Volume 31:Number 7(2021)
• Page Start: 531
• Page End: 537
• Publication Date: 2021-09-02
• Subjects: Cryopreservation -- human peripheral blood leukocytes (HPBL) -- human lung-derived embryonic fibroblast (FLECH-104) -- alkaline comet assay -- highly damaged cells; Analytical toxicology -- Periodicals; Toxicology -- Periodicals; Toxicology -- Methodology -- Periodicals; 615.907
• DOI: 10.1080/15376516.2021.1933286
• Languages: English
• ISSNs: 1537-6516

Abstract

Abstract: Single-cell gel electrophoresis (comet assay) is a valuable test that can be used in ecotoxicological, epidemiological, and biomonitoring contexts. We assessed the effects of short- (without cryopreservation) and long-term (with cryopreservation) storage of DMEM-cultivated human peripheral blood leukocytes (HPBLs) and a human lung fibroblast cell line (FLECH-104) on comet assay results. Samples were stored for 6 or 24 h at room temperature (23°С) or 4 °C and frozen at −80 °C or −196 °C for 1, 2, or 4 weeks. Short-term storage led to significant increases in the comet tail intensity (TI) and Olive tail moment (OTM) in HPBL and FLECH-104 samples. Freezing FLECH-104 samples at −80°С and −196°С resulted in TI mean increases, with no differences in OTM. All frozen HPBL samples did not exhibit significant increases in TI or OTM, and instead exhibited a slight decrease in TI versus the control at both −80 °C and −196 °C. Increased frequency of highly damaged cells was observed in FLECH-104 and HPBL cultures during both short-term storage and after freezing, which may indicate a significant destructive effect. Therefore, freezing of cell cultures and whole blood according to our protocol is not recommended.