NDLI: The Cryopreservation of a Tissue Engineered Dermal Replacement by Programmed Freezing

Content Provider	IEEE Xplore Digital Library
Author	Wang, X. Hua, T.C. Yang, G.H. Cui, L. Liu, W. Cao, Y.L.
Copyright Year	2005
Description	Author affiliation: Inst. of Thermal Sci. in Medicine, Shanghai Univ. of Sci. & Technol. (Wang, X.; Hua, T.C.)
Abstract	Long-term storage of engineered bio-artificial tissues is required to ensure the off-the-shelf availability to clinicians due to their long production cycle. Cryopreservation is likely the choice for long-term preservation. This study investigated the effects of dimethyl sulfoxide (DMSO) concentrations, cooling rates, cryoprotectant medium treatment methods and seeding on the cell viability of a tissue-engineered dermal substitute. The dermal fibroblast was cultured on a polyglycolic acid (PGA) scaffolding at 37degree C and a 5% $CO_{2}$ atmosphere for 14 days, and dermal slices (10times3times1.5 mm) were used in the experiment. Two groups were defined - fresh control and experimental group. The freezing was carried out in a computer-controlled programmable freezer at 0.5 degree C/min, 1degree C /min and 2degree C /min in 1.4M, 2.1M and 2.8M DMSO from 4degree C to -60degree C and then plunged into the liquid nitrogen tank immediately. After 24 h of cryopreservation, the samples were removed from liquid nitrogen and placed at room temperature in air for 0.5-1 min, prior to submergence in a water bath at 37degree C . Viability of fresh control and cryopreserved dermal slices were determined with our modification of the MTT (3-[4,5-Dimethythiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. The effects of two cryoprotectant medium treatment methods and seeding (at -6degree C ) on cell viability were also assessed. A cryopreservation protocol of 1degree C /min in 1.4M DMSO from 4degree C to -60degree C , and seeding at -7degree C proved optimal for this tissue-engineered dermal replacement, provided the dermal slice was transferred into the cryopreservation bag containing 1mL 1.4M DMSO cryoprotectant medium, which was kept at 4degree C , seal the bag and hold for 15 min at 4degree C prior to freezing. The viability of the dermal replacement treated by this protocol was maintained 80% of the fresh control viability. And the micrograph confirmed that after 48 h incubation following thawing, the dermal fibroblast can regain its natural shape and functions. No fracture was observed
Starting Page	494
Ending Page	497
File Size	289535
Page Count	4
File Format	PDF
ISBN	0780387414
DOI	10.1109/IEMBS.2005.1616455
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2006-01-17
Publisher Place	China
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Dermis Fibroblasts Nitrogen Protocols Production Cooling Electronics packaging Atmosphere Temperature Seals
Content Type	Text
Resource Type	Article

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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