RE: (Fwd) sorting safety

Hi Kent:
We recently went through this painstaking process in deciding how best to
handle sorting of primary human tissue, and in our case samples are
untested, mainly because our facility is only approved at the BSL2 level.
There was quite a bit of discussion surrounding the testing part (you can
search the archives for my postings) since it was felt that testing for a
few pathogens may only provide a false sense of safety. It was decided to
follow the OSHA guidelines which basically say you should assume all samples
are infected. Well this is what opened the can of worms since we are
essentially when we sort we are aerosolizing samples which we are assuming
to be infected. The question arose as to whether these samples should be
considered BSL3 once introduced to sorting. We could have gone round and
round forever discussing this point, but the bottom line was to make sure
the sort operators were as safe as we could make them and that they were
comfortable with the protection available. We ended up sorting these samples
in our BSL2 approved facility under BSL2+ precautions. We have baseline
serum samples drawn for those operators performing these sorts. We have a
Cytek aerosol containment hood installed on our FACSVantage SE sorter which
we have validated through the use of a fine particle measuring device (done
through our EHS) and Glow Germ particles. In addition we have invested in a
PAPR Hood system which the operator wears in addition to the aerosol
containment device. Gowns and gloves are worn at all times and the room is
closed off to all except the operator during the sort, unless others are
also appropriate equipped with a PAPR device. A short Biosafety
Questionnaire is required to be completed for each and every sample to be
sorted and must be signed by the PI. The sorter and surrounding lab space is
decontaminated with 10% bleach following the completion of the sort. Some
may think this is overkill, but better to be safe than sorry. I don't agree
with the argument that because there is no documented case of an infected
sort operator that this is justification that we do not need to worry. I am
not aware of any formalized program that tests sort operators on a regular
basis. Infection and disease are two very different things. Some viral
pathogens to which we are potentially exposed may be in fact more of a risk
for cancer than infectious disease. There are lots of different infiltrating
cells within a tumor that could potentially harbor pathogens.
My personal opinion, for what it is worth, is that all primary human tissue
should be sorted in a BSL3 facility using BLS3 practices. If you search the
archives you will see this has been discussed in detail before. There is
also a summary of responses that Paul Robinson put together on the Purdue
CD8 which I believe you can get from Bangs Labs. I also believe that the
Biosafety Committee of ISAC will be generating some type of recommendations
for sorting primary human tissue in the near future. Perhaps Steve Perfetto
or someone else from the committee might want to jump into the discussion. 

Joanne Lannigan, MS
Director, Flow Cytometry Core Facility
University of Virginia
Jordan Hall, Room 7067
P.O. Box 800734
Charlottesville, VA 22908-0734
Office: 434-924-0274
Lab: 434-243-2695
Fax: 434-982-1071
email: joannelannigan@virginia.edu

> -----Original Message-----
> From: kclaypool@mdanderson.org [mailto:kclaypool@mdanderson.org]
> Sent: Friday, April 22, 2005 1:13 PM
> To: Cytometry Mailing List
> Subject: (Fwd) sorting safety
> 
> 
> Dear Flow Community,
> 
> Does your Flow Cytometry facility sort live (unfixed) clinical (tested)
> patient samples? Please write me with contact information so that I
> may inquire about your safety policies and procedures.
> 
> Specifically we have an investigator that would like to sort primary
> tumor (human: prostate, glioblastoma, cervical, etc.) stem cells that
> have been clinically tested for HIV, HBV, etc. Any feedback on the
> potential operator risks with pluripotent cancer stem cells? Minimum
> facility requirements? BSL1, class 1000 lab or clean room, etc?
> 
> Thanks!
> 
> Kent Claypool
> Department of Carcinogenesis
> University of Texas M.D. Anderson Cancer Center
> Science Park - Research Division
> 1808 Park Road 1C
> P.O. Box 389
> Smithville, TX 78957
> 
> Tel. 512-237-9427
> Fax: 512-237-9589