Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet.
BACKGROUND
The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2.


METHODS
After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions.


RESULTS
The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabinet, does not influence the conditions for side stream stability or the analytical precision of the FACS Vantage cell sorting system. In addition, tests performed to assess aerosol containment during operation of the special biosafety cabinet demonstrated that the cabinet-integrated FACS Vantage unit (CIF) satisfies the conditions for class II cabinets. In the context of gene transfer experiments, the CIF facility was used to sort hematopoietic progenitor cells under biosafety level 2 conditions.


CONCLUSIONS
The newly designed biosafety cabinet offers a practical modality for improving biosafety for operators and samples during cell sorting procedures. It can thus also be used for sorting experiments with genetically modified organisms in conformance with current biosafety regulations and guidelines.
Institute of Cell Biology (Cancer Research), West German Cancer Center Essen, University of Duisburg-Essen, Medical School, Essen, Germany.
Klaus
K
Lennartz
Mengji
M
Lu
Michael
M
Flasshove
Thomas
T
Moritz
Uwe
U
Kirstein
D000336:Aerosols; D000392:Air Movements; D018952:Antigens, CD34 / Q000032:analysis; D002469:Cell Separation; D003264:Containment of Biohazards / Q000295:instrumentation*; D003564:Cytidine Deaminase / Q000096:biosynthesis / Q000235:genetics; D004780:Environment, Controlled; D004866:Equipment Contamination; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics; D017408:Guidelines as Topic; D015386:Hazardous Substances; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism / Q000821:virology; D006801:Humans; D016272:Occupational Health; D030781:Organisms, Genetically Modified; D012190:Retroviridae / Q000235:genetics; D017751:Safety Management / Q000379:methods*; D014161:Transduction, Genetic
D000336:Aerosols; D018952:Antigens, CD34; D015386:Hazardous Substances; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D003564:Cytidine Deaminase