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Cryopreservation of cellular products in a closed‐bag system with an incorporated dimethyl sulfoxide–resistant sterile filter outside of cleanroom facilities
Author(s) -
Humpe Andreas,
Jansen Pal,
Tischer Bernd Karsten,
Schubert Sabine,
Beck Christian,
Adamzik IlseDorothea,
Maas JensHolger,
Strate Alexander,
Gramatzki Martin,
Riggert Joachim
Publication year - 2007
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2007.01232.x
Subject(s) - cryopreservation , dimethyl sulfoxide , leukapheresis , cleanroom , andrology , biology , chemistry , medicine , materials science , stem cell , microbiology and biotechnology , cd34 , nanotechnology , embryo , organic chemistry
BACKGROUND: Manipulations, for example, cryopreservation, of cellular therapeutics carried out in an open system must be performed in a class A environment with surrounding class B environment. To avoid cleanroom facilities, a new closed‐bag system with an incorporated dimethyl sulfoxide–resistant sterile filter for cryopreservation of cellular products was evaluated at two different centers. STUDY DESIGN AND METHODS: A total of 44 different products (22 buffy coats [BCs] and 22 leukapheresis [LK] products) were split and cryopreserved in parallel in cleanroom facilities (Method I) and with the closed system on the bench of a “normal” laboratory (Method II). Viability analyzed by 7‐aminoactinomycin D staining and flow cytometric analysis and sterility of the products were analyzed. RESULTS: Independent of the cellular source (BC or LK), the median viability of CD45+ cells decreased significantly (p < 0.01) during cryopreservation: namely, in BCs, −15.8 percent with both methods, and in LK products, −5.4 percent with Method I and −4.8 percent with Method II, respectively. CD3+ as well as CD14+ cells exhibited a similar pattern and were also found significantly (p < 0.01) diminished after thawing independent of the handling system. For CD19+ cells, the small decrease of viability was only for the BC group significant (p = 0.027) when the cells had been processed with Method I. No bacterial contamination was detected neither in fresh products nor in products after cryopreservation. CONCLUSION: The closed system for cryopreservation of cellular products appears to be equivalent to cleanroom‐based methods regarding cellular integrity and sterility when appropriate quality of sterile filters is assured.