A simple tube adapter to expedite and automate thawing of viably frozen cells.
Although cryopreserved cell specimens are used throughout biomedical research, the process for thawing samples is labor-intensive and prone to error. Here we describe a small laboratory device that couples an uncapped vial of frozen cells to a conical tube containing warm cell culture media. The entire complex is loaded directly into a centrifuge; within 5min, cells are thawed and diluted out of toxic cryopreservation medium. The recovery and viability of cells are slightly reduced compared to the common (traditional) method. However, antigen-specific T-cell function is not affected. Since no technician time is required (beyond uncapping of vials), our device allows the parallel processing of as many samples as a centrifuge can hold (up to 96, in some models). Moreover, since the samples are not thawed manually in a water bath, the problems associated with technician-to-technician differences in sample handling are minimized, as is the potential for contamination. Importantly, the elimination of substantial labor involving subjective decisions standardizes this process and can reduce variability in results from cryopreserved specimens.
ImmunoTechnology Section, Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, United States.
Margaret
M
Beddall
ImmunoTechnology Section, Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, United States. Electronic address: pchattop@mail.nih.gov.
Pratip K
PK
Chattopadhyay
ImmunoTechnology Section, Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, United States.
Shing-Fen
SF
Kao
ImmunoTechnology Section, Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, United States.
Kathy
K
Foulds
ImmunoTechnology Section, Vaccine Research Center, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, United States.
Mario
M
Roederer
D057205:Automation, Laboratory; D002470:Cell Survival; D002498:Centrifugation / Q000295:instrumentation* / Q000592:standards; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000633:toxicity; D003470:Culture Media / Q000737:chemistry; D004867:Equipment Design; D005434:Flow Cytometry; D006801:Humans; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
D003451:Cryoprotective Agents; D003470:Culture Media
Cryopreservation; Immunoassay; T-cells; flow cytometry