Handling Human Cell Lines

Human Cell Lines Human cell lines, either primary or established lines, are commonly used in biomedical research. Both primary and established cell lines need to be handled in accordance with the OSHA Bloodborne Pathogens Standard and require biosafety level 2 procedures. The Radiological & Environmental Management department (REM) is responsible for the University Bloodborne Pathogen and Biosafety programs.

Training Requirements Users of primary cell lines, those obtained directly from a person or blood bank, must attend annual Bloodborne Pathogen training. Contact REM to schedule. Users of established cell lines, those obtained from a commercial source, must review this Bloodborne Pathogen Power Point.

Regulations and Safety Manuals The Occupational Safety and Health Administration (OSHA) requires all employees with occupational exposure to bloodborne pathogens to participate in an annual training program (29 CFR 1910. 1030). Purdue University Bloodborne Pathogens Exposure Control Plan Purdue University Biological Safety Manual

Bloodborne Diseases - HIV Human Immunodeficiency Virus (HIV) Incubation period 1 –3 months for the detection of antibodies. Symptoms depend on the degree of immunodeficiency. HIV progresses into AIDS which has a 100% fatality. There is no vaccine available. Generally, individuals acquire many different types opportunistic infections which result in anorexia, chronic diarrhea, weight loss, fever and fatigue. HIV is transmitted through exposure to contaminated human blood, blood products, or tissue. Occupational exposures include sharps accidents (needles or glass), splash, and abrasive contact.

Bloodborne Diseases - HBV Hepatitis B Virus (HBV) Incubation period 8 – 12 weeks average for onset. Symptoms range from the undetectable (carriers) to fatal liver function failures. HBV is transmitted through exposure to contaminated human blood, blood products, or tissue. Occupational exposures include sharps accidents (needles or glass), splash, and abrasive contact. A vaccine is available.

Bloodborne Diseases - HCV Hepatitis C Virus (HCV) Incubation period 6 – 9 weeks average for onset. Chronic infection may persist for 20 years. Symptoms range from the undetectable (carriers) to fatal liver function failures. HCV is transmitted through exposure to contaminated Human blood, blood products, or tissue. Occupational exposures include sharps accidents (needles or glass), splash, and abrasive contact. There is no vaccine available.

Precautions Job tasks that potentially expose employees to human cell lines should be documented and discussed before work or contact takes place. Practice Standard Precautions; the concept that all blood products, cell lines, or tissue are contaminated with pathogens and must be handled with protective safeguards.

Reducing Exposure Reduction of human cell line exposure includes: Applying engineering controls like sharp containers, Enclosing equipment, using barriers that isolate the employee Using Personal Protective Equipment (PPE) outerwear such as gloves, gowns, and face shields that prevent contact with blood products. Changing work practice behaviors where human cell lines are handled

Personal Protective Equipment (PPE) PPE is the last barrier of defense between the user and the hazard. PPE must be available for every body part as well as respiratory protection. Proper selection of PPE is critical for each individual. Know how to don and doff each piece of PPE. A typical selection of PPE for biohazard application would be the use of nitrile or latex gloves, goggles, lab coat, closed toe shoes, and perhaps a N-95 dust and vapor mask.

Sharps and Waste Contaminated sharps such as broken glass, razor blades, or needles must be placed in properly biohazard labeled puncture proof container. Contaminated PPE must be placed in properly labeled water impervious bags. Both of these contaminated waste materials must be decontaminated before disposal. Decontaminate by autoclaving, incineration, or soaking in bleach. Sharps Procedure Sharps and Infectious Waste: Handling and Disposal

Sharps Handling Sharps are items that are capable of puncturing, cutting, or abrading the skin, i.e., broken plastic or broken glassware, glass or plastic pipettes, scalpels, razor blades, needles, hypodermic needles, etc… Do not place any sharps into the regular trash. Needles and razor blades must be disposed of in puncture proof plastic containers. Clean broken glass should be collected in a cardboard box. When ready for disposal, tape the box shut and label it “SHARP OBJECTS/GLASS - DISCARD”. It is prudent to affix a “safe for disposal” sticker to the box as well.

Waste Handling - Biohazard Bags BIO-RELATED WASTE MATERIALS: Dispose of in properly labeled water impervious autoclavable bags (usually red or orange bags). Do not over fill bags. Utilize proper personal protective equipment and wash hands after handling. Keep bags closed until they are ready to be decontaminated. Do not dispose of a bio waste bag in the regular trash. Do not place these bags in public areas such as hallways. Contact REM for removal of decontaminated and autoclaved materials.

Commonly Used Disinfectants Know what disinfectant to use for the specific bio-agent being handled. Alcohols: ethyl or isopropyl alcohol at 70-80 percent concentration is a good general purpose disinfectant; not effective against bacterial spores. Phenols: Effective against gram-positive bacteria, fungi, and lipid-containing viruses. Quaternary Ammonium Compounds: Cationic detergents which are strongly surface active; extremely effective against lipophilic viruses; not effective against bacterial spores. Chlorine: Low concentration (50-500 ppm) active against vegetative bacteria and most viruses; higher concentration (2500 ppm) required for bacterial spores; corrosive to metal surfaces; must be prepared fresh; laundry bleach (5.25 percent chlorine) may be diluted and used as a disinfectant. Iodine: Recommended for general use; effective against vegetative bacteria and viruses; poor activity against bacterial spores. Many of these disinfecting agents can be irritating and toxic and should be used in accordance with label direction for personal protective equipment, concentration, and contact time.

Vaccinations Hepatitis B vaccine inoculations are available to employees exposed to human blood products and cell lines at no cost. There are a total of three scheduled injections spanning six months, followed by titer blood work. Adverse reactions occur in 1 - 10% of injections. Symptoms could include fever, headache, and dizziness. Effective immunity is usually permanent, although individuals in very high risk groups should have boosters every five years. Contact REM to initiate the vaccination process.

Accidental Exposure Exposure is defined as a cut, puncture, or splash into the eyes with contaminated sharps or cell line materials. In the event of an accidental exposure the employee should wash the exposure site with soap and water, inform their supervisor immediately, and seek medical attention; be sure to inform the health care provider that you were exposed to a human fluid or cell line.

Reporting The supervisor is required to notify REM so that a follow-up investigation can be conducted. PUSH will arrange for the source blood to be tested for HIV/HBV and HCV. The results of this blood testing will be confidential and relayed to the exposed employee as soon as possible.

Disease Statistics Links Indiana State Department of Health (ISDH) Indiana Epidemiology Newsletters Guidelines for the Management of Hepatitis C Currently Living in Indiana with HIV Disease by Last Known County of Residence by Mode of Transmission Department of Health and Human Services - Center for Disease Control and Prevention (CDC) National Center for HIV/AIDS, STD & TB Prevention

Contact Information Robert Golden, Biological Safety Officer, rwgolden@purdue.edu, 41496 Office of the Vice President for Research Radiological & Environmental Management

Training Verification Click here to print the Handling Human Cell Lines Training Record. Print the form, complete the required information, send a copy to R. Golden/REM/CIVL, and give the original to your PI, supervisor, or instructor to place and retain in the departmental safety training files.