Not wearing gloves during tissue culture - (Feb/18/2013 )

Hi all,

I have a problem in my lab in that my PI does not wear gloves for tissue culture AND she trains all the BS and MS students in her lab (of which there are MANY) not to wear gloves either. I've been working in biology since college and to me wearing gloves is not optional - I always wear gloves and and very careful with my sterile technique. But now we have an upcoming Biosafety meeting for the department, which I feel is a complete farce before it begins, when she doesn't even have people wearing gloves. For instance, some cancer cell lines are viral-transformed and may still emit viral particles and/or are BSL2. I also don't like the students coming into the tissue culture hoods and not wearing gloves, therefore contaminating everything in the hood with their hands (mycoplasma and other contaminants).

I need some information to confront the PI with to change this policy as I feel not wearing gloves is very bad, for many reasons.

So I would like to know from other biologists, have you ever worked in a lab where you could do tissue culture without gloves?

And how would you react to seeing someone not wearing gloves while doing tissue culture?

Wow, tissue culture without gloves. That said, it could be a generational thing - scientists 30-40 years ago mouth-pipetted, used bare hands to handle stuff etc. But with tissue culture without the appropriate safety-wear is appalling. There will be an increased risk to contaminate your cell lines with microbes (mycoplasma, bacteria, yeast, fungi...) and you also run the risk of getting some unwanted stuff from the cell/media/reagents/chemicals.

Do you have a department or person who deals with health & safety issues and/or biosafety? If so, I would consult them regarding your PI.

yea she trained in the 80s back with mouth pipetting and all that, but to this day she insists it's BETTER to not use gloves... unfortunately our small college has a completely ineffective safety officer, but I am trying to gang up on my PI with the department's "jack of all trades" tech, but she has a lot on her plate. So really I just want to get some outside opinions about it to add to my own reasons to present to her before our lol-biosafety meeting.

btw according to the CDC they say you must wear gloves even for BSL1 work: http://www.cdc.gov/biosafety/publications/bmbl5/bmbl5_sect_iv.pdf

Another point is that if they don't wear gloves and touch other things in the lab, virtually everything might be contaminated with whatever they took up from the cell culture...
Don't understand why it should be better not to wear gloves - of course there are these people who touch everything else with their gloves too and think thats sterile just because they have gloves, but as long as you disinfect gloves after touching something outside the hood all is fine...
Do they use gloves for PCR pipetting?

Dear Shirohands,

When I first did cell culture in the late 1970's we:

Used glass pipettes and flasks
We used a bunsen burner to flame all the pipettes and flask tops to ensure sterility.
We used lots of 70% Industrial methylated spirits (IMS) to clean surfaces and flasks.
All of this was done on a normal bench......i.e. no laminar flow cabinets or class II cabinets

WE DID NOT USE GLOVES AS THIS WAS SEEN AS A HAZARD WHEN USING IMS AND A FLAMING BUNSEN BURNER. THE BURNS WHEN HAVING AN ACCIDENT WITH GLOVES ON (WHEN USING A BUNSEN AND IMS) WERE SEEN TO BE ALOT MORE DAMAGING.


We then moved onto using laminar flows, but generally speaking these were horizontal laminar flows which protected the cells from our possible contamination. Again we were still using IMS and no gloves.

In the late 1980's Class II cabinets came in to give operator protection as well as product protection. Plastic single use pipettes and flask came in so there was no longer a need to use a bunsen burner. Technically speaking it is difficult to get class II cabinets to pass their KI and containment tests with a bunsen burner.

BECAUSE OF THIS AND THE INCREASE IN USING TRANSFORMED CELLS RATHER THAM PRIMARIES, IT BECAME COMMON PRACTICE TO WEAR GLOVES WHEN MANIPULATING CELLS IN CLASS II CABINETS. THE RISK OF HAVING A FIRE DUE TO USING BUNSENS AND IMS HAD BEEN REMOVED.

I train young people in the "dark art" of cell and tissue culture in the University I work in. I have over 35 years experience in cell and tissue culture and I recommend using gloves AT ALL TIMES. It is a barrier to all the bacteria, fungi and mycoplasma's that are living on our hands at all times.

And to be factual, mouth pipetting finished in the late 1970's. In the pharmacuetical company I worked in from 1977-1995, I never once witnessed mouth pipetting.

Hope this is useful and give you some facts to back up your arguement. Our health and safety officer in new staff inductions always recommends wearing gloves for cell and tissue culture.

Kindest regards

Uncle Rhombus

Well the issue has been resolved but it wasn't effortless... I was not budging on it for safety reasons, and after much arguing she agreed to comply for legal reasons.

According to what I found, I believe that if someone wanted to sue her or the institution for not wearing gloves with tissue culture, they could. To be in line with current Federal regulations, it is probably illegal to not wear gloves during tissue culture. btw here is the legal justification, according to what I found:


Here is where the current CDC BSL Guidelines are:
http://www.cdc.gov/biosafety/publications/bmbl5/bmbl5_sect_iv.pdf

Which states, for both BSL1 and BSL2: "Gloves must be worn to protect hands from exposure to hazardous materials."

Here is the ATCC MSDS for cell lines:
https://www.atcc.org/~/media/DA6CC2AA0A034BD998F2F069C6552299.ashx

which states: "Various animal Cell Cultures at Biosafety Level 1 or 2 ... this product should be handle according to good lab practices, with proper personal protective equipment...Handle as a potentially biohazardous material...Universal Precautions according to 29 CFR 1910.1030 should be followed at all times when manipulating these cell lines."

29 CFR 1910.1030 are the Federal regulations concerning Bloodborne pathogens, found online at:

http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_id=10051

Per federal US Dept. of Labor OSHA Regulations 29 CFR 1910.1030 (d)(3)(ii):

"The employer shall ensure that the employee uses appropriate personal protective equipment."

Per OSHA Regulations 29 CFR 1910.1030 (d)(3)(ix):

"Gloves shall be worn when it can be reasonably anticipated that the employee may have hand contact with potentially infectious materials."

So in the end, even if you consider something to be harmless enough to get on your skin, it must be treated as a POTENTIAL hazard no matter what. The key word being "potentially" which as I see it is valid for all BSL1 and 2 organisms, and I also gave the reasoning that cancer cell lines derived from humans should still be treated as a bodily fluid, whether or not any real danger exists. Anyway, there are a lot of other reasons to wear gloves of course, but we are finally in agreement here !

Of course she has entirely different tissue culture technique than I do and doesn't want me regulating her on that, which I will not, because probably no one will sue you about how you split your cells, but it's possible they could sue you over not wearing gloves.