Elements and Performance Criteria

The range of variables relates to the unit of competency as a whole. It allows for different work environments and situations that will affect performance.
Where reference is made to industry Codes of Practice, and/or Australian/international standards, it is expected the latest version will be used.

All work is performed according to the legal/regulatory framework of regulations, codes and guidelines including
AS ISO/IEC 17025 General requirements for the competence of testing and calibration laboratories
ISO 9000 series Quality management and quality assurance standards
AS 1678 Emergency procedures guide for hazardous materials
AS 2243 Safety in laboratories
AS 2243.3 Microbiology laboratories
AS 2243.8 Fume hoods
AS 2252 Biological safety cabinets
AS 2982 Hand washing facilities
SAA HB9 Occupational personal protection, and other relevant standards for protective, clothing (for example, AS 2161, AS 2210, AS 1337 and AS 1338)
AS 4187 Code of Practice for cleaning, disinfecting and sterilising reusable medical and surgical instruments and equipment, and maintenance of associated environments in health care facilities.
guidelines for the operation of classes of laboratories
Office of the Gene Technology Regulator (OGTR) guidelines for working with genetically altered organisms
Good Manufacturing Practice (GMP)
Good Laboratory Practice (GLP)
Therapeutic Goods Act.

Enterprise procedures may include
standard operating procedures (SOPs)
quality assurance procedures
verified test methods
laboratory manuals.

Hazards may include
biohazards, such as infectious agents, oncogenic DNA
chemical and radiation hazards
allergenic factors
cryogenic liquids, such as nitrogen
heat from burners, molten agar
ultraviolet light
sharps
contaminated clothing.

Hazard control measures may include
ensuring access to service shut off points
recognising and observing hazard warnings and safety signs
labelling of samples, reagents, aliquoted samples and hazardous materials
handling and storage of hazardous materials and equipment in accordance with labelling, materials safety data sheets and manufacturer's instructions
identifying and reporting operating problems or equipment malfunctions
cleaning and decontaminating equipment and work areas regularly using enterprise procedures
using personal protective clothing and equipment, such as gloves, safety glasses, coveralls, gown, body suits, respirators
using containment facilities (PCII, PCIII and PCIV physical containment laboratories), containment equipment (biohazard containers, laminar flow cabinets, Class I, II and III biohazard cabinets) and containment procedures
reporting abnormal emissions, discharges and airborne contaminants, such as noise, light, solids, liquids, water/waste water, gases, smoke, vapour, fumes, odour and particulates to appropriate personnel.

Tissue culture equipment and facilities may include
growth cabinets
culture vessels, growth chambers, sterile containers, culture plates, flasks and bottles
autoclaves
positive filtration apparatus
auto pipettes and pipette pumps
cell counting chambers
incubators, including specialised atmosphere, carbon dioxide
binocular inverted microscope
centrifuges
cryogenic vessels and transfer equipment, liquid nitrogen.
Selection criteria for media, materials and equipment may include costs, ease of cleaning or sterilisation, maintenance of cell growth

Pre-use checks include
performing routine maintenance
checks on raw materials and consumables include use by date, possible contamination and storage conditions.

Cells and tissues may include
animal cell lines, such as hybridoma, liver, epidermal, lymphoblastic, fibroblastic
plant cell lines, such as tobacco, arabidopsis, soya bean, tomato, roses, meristomatic tissue
yeasts
fungi
sperm, ova and embryos
adherent and suspension cultures.

Preparing a primary culture may include
thawing of cryopreserved cells and monitoring of cell recovery
enzymatic disaggregation from tissue
mechanical disaggregation from tissue
primary explant technique
pre treatment
selection techniques, such as cloning, micromanipluation, use of selective media, density gradient centrifugation, selective adhesion techniques and selective detachment.

Monitoring growth of tissue and cell lines may include
identification of normal and abnormal cells viewed using an inverted stereo microscope
recognition of contamination by cytopathic changes to cells, biochemical tests, gene detection and microbiological culture
testing for products, such as insulin
checking growth rates
performing viable cell counts, such as the dye exclusion test, Trypan Blue viability stain to determine percentage viability and total cell concentration
staining and assessment of morphology eg by Giemsa
karyotype analysis.
Preservation of cell lines can include freezing, cryopreservation (dry ice, liquid nitrogen).

Records may involve
paper or Laboratory Information Management Systems (LIMS)
cataloguing of all cell lines
stock levels
viability test results.

Health, safety and environment
All operations to which this unit applies are subject to stringent health, safety and environmental (HSE) requirements, which may be imposed through State or Federal legislation, and these must not be compromised at any time. Where there is an apparent conflict between performance criteria and HSE requirements, the HSE requirements take precedence.
All operations assume the potentially hazardous nature of samples and require standard precautions to be applied. Users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council and State and Territory Departments of Health. All operations are performed in accordance with standard operating procedures.

The Evidence Guide describes the underpinning knowledge and skills that must be demonstrated to prove competence.
Critical aspects of competency

Competency must be demonstrated in the ability to perform consistently at the required standard. In particular, assessors should look to see that the candidate:

works safely and satisfies all legal and regulatory requirements, including the use and care of safety cabinets
demonstrates chain of custody for all cells, cell lines and tissues
prepares, dilutes and sterilises reagents and culture media that are fit for purpose
chooses and justifies appropriate media and substrate material based on cost, cleaning, sterilising and maintenance of cell growth
successfully passages cell cultures by subculturing
grows cell lines and tissue to specifications without contaminating the original sample and the environment
counts cells, identifies a wide range of cell types and contaminants and recognises normal and abnormal cells
monitors cell growth and recognises and troubleshoots problems, such as contamination
stores cells so that they remain viable
maintains accurate, traceable records of cell lines and tissues and logs of procedures and work completed.

Underpinning knowledge

Competency includes the ability to apply and explain:

purposes of cell lines
normal and abnormal cell morphology
terminology, such as: cell lines, growth media, primary culture, passaging, passage number, subculture, anchorage dependent cells, suspension culture, monolayer, confluent, cell line, cell strain, contact inhibition, diploid, viability
cell biology - structure, physiology, function, physiological cell growth requirements, nutrient requirements, respiration, temperature, growth cycle
critical components of the cell environment and their effects on cell growth, such as: pH, temperature, buffering, osmotic pressure, osmolarity, viscosity and foaming
types of tissue used as source material, such as: embryonic, adult or malignant tissue
techniques for characterising a cell line
the differences between finite and continuous cell lines
characteristics of cell culture media and substrates
nature of the substrate (for example, solid, semi-solid, gel or sponge, glass, disposable plastics, three dimensional matrices)
techniques for pre treating substrates (for example, feeder layers, chemical treatments, such as poly D-lysine, collagen, gelatine and fibronectin)
role of ingredients in media - (for example, salts, carbohydrates, amino acids, vitamins, hormones, growth factors, serum, antibiotics)
contaminants, such as: endotoxins, bacteria, yeast, fungi and Mycoplasma
requirements, typical problems and procedures associated with the production of specific cell lines
relevant health, safety and environment requirements.

Assessment context and methods
This unit of competency is to be assessed in the workplace or simulated workplace environment.

The following assessment methods are suggested:

review of records of cell lines and tissues produced by the candidate
periodic observation of the candidate establishing and maintaining viable cell lines
feedback from peers and supervisors to confirm that workplace procedures are consistently followed and that results meet workplace requirements
oral and/or written questioning.
In all cases, practical assessment should be supported by questions to assess underpinning knowledge and those aspects of competency which are difficult to assess directly. Questioning techniques should suit the language and literacy levels of the candidate.

Interdependent assessment of unit

This unit of competency may be assessed with:

PMLMAIN300B Maintain the laboratory fit for purpose.

Resource implications

Resources may include:

laboratory equipped with appropriate test equipment/instruments, standards and reagents
enterprise procedures and standard methods
relevant tissues and cell lines.

This competency in practice
A laboratory technical officer works at a research institute that genetically modifies myocardial cell lines to express Angiotensin II receptors and modify their action. Their role in the team is to grow the cells. This involves selecting the appropriate media, growth conditions and equipment and carefully monitoring cell growth. Each day, they visually check the cells and, when necessary, modify pH, temperature, buffering, osmolarity and substrates to enhance growth. The technical officer keeps accurate and legible records of cells, cell lines, tissues, observations and details of all modifications so that the team has a complete, reliable record of all work done.
A laboratory technical officer works at a metropolitan pathology laboratory. Their role is to prepare and use cell cultures for the initial isolation of viruses, such as the herpes simplex (HSV I and II). They routinely subculture human embryonic lung (HEL) cells using appropriate media, flasks and aseptic techniques in a Class II biohazard cabinet. They inoculate each flask with 0.1mL of patient swab washings and incubate them at 37C for seven days. They also use appropriate positive and negative controls as required by the laboratory's quality assurance procedures. Each day, the technical officer examines the cell monolayer for distinctive changes (cytopathic effect). When the effect is detected, they seek confirmation of the changes from a senior technician. The flask is then sent for immunofluorescent testing to identify the virus isolate.

The seven key competencies represent generic skills considered for effective work participation. The bracketed numbering against each of the key competencies indicates the performance level required in this unit. These are stand-alone levels and do not correspond to levels in the Australian Qualifications Framework (AQF).
Level (1) represents the competence to undertake tasks effectively
Level (2) represents the competence to manage tasks
Level (3) represents the competence to use concepts for evaluating and reshaping tasks.
Communicating ideas and information
(2)

Collecting analysing and organising information
(3)

Planning and organising activities
(2)

Working with others and in teams
(2)

Using mathematical ideas and techniques
(2)

Solving problems
(3)

Using technology
(2)