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Evidence Guide: MSL975013A - Perform tissue and cell culture techniques

Student: __________________________________________________

Signature: _________________________________________________

Tips for gathering evidence to demonstrate your skills

The important thing to remember when gathering evidence is that the more evidence the better - that is, the more evidence you gather to demonstrate your skills, the more confident an assessor can be that you have learned the skills not just at one point in time, but are continuing to apply and develop those skills (as opposed to just learning for the test!). Furthermore, one piece of evidence that you collect will not usualy demonstrate all the required criteria for a unit of competency, whereas multiple overlapping pieces of evidence will usually do the trick!

From the Wiki University

 

MSL975013A - Perform tissue and cell culture techniques

What evidence can you provide to prove your understanding of each of the following citeria?

Interpret and schedule production requirements

  1. Review client request and confirm quantity and nature of cells, tissue or products
  2. Select, appropriate media, materials, equipment and methods
  3. Plan parallel work sequences to optimise production
  4. Maintain a chain of custody, traceable to the worker, for all cells and tissues
Review client request and confirm quantity and nature of cells, tissue or products

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Select, appropriate media, materials, equipment and methods

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Plan parallel work sequences to optimise production

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain a chain of custody, traceable to the worker, for all cells and tissues

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Work safely according to the legal and regulatory framework

  1. Ensure work practices and personal actions conform to regulations, codes, guidelines and enterprise quality assurance procedures
  2. Identify hazards and enterprise control measures associated with the sample, preparation methods, reagents and equipment
  3. Select, fit and use personal protective clothing and safety equipment
  4. Address hazards and incidents as they arise
  5. Ensure the safe disposal of biohazardous materials and other laboratory wastes
Ensure work practices and personal actions conform to regulations, codes, guidelines and enterprise quality assurance procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Identify hazards and enterprise control measures associated with the sample, preparation methods, reagents and equipment

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Select, fit and use personal protective clothing and safety equipment

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Address hazards and incidents as they arise

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure the safe disposal of biohazardous materials and other laboratory wastes

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Assemble and maintain tissue culture equipment

  1. Assemble, sterilise or decontaminate equipment according to enterprise procedures
  2. Perform pre-use and safety checks in accordance with relevant enterprise and operating procedures
  3. Identify faulty or unsafe components and equipment and report to appropriate personnel
  4. Decontaminate area and equipment after use
Assemble, sterilise or decontaminate equipment according to enterprise procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Perform pre-use and safety checks in accordance with relevant enterprise and operating procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Identify faulty or unsafe components and equipment and report to appropriate personnel

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Decontaminate area and equipment after use

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare and test cell and tissue culture media

  1. Confirm media specifications and processes/methods
  2. Prepare culture media to suit client request
  3. Sterilise culture media and check for sterility
  4. Perform quality control checks to ensure that culture media is fit for purpose
  5. Store culture media in accordance with specifications
Confirm media specifications and processes/methods

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare culture media to suit client request

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Sterilise culture media and check for sterility

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Perform quality control checks to ensure that culture media is fit for purpose

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Store culture media in accordance with specifications

Completed
Date:

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Obtain, monitor and maintain tissue and cell lines

  1. Retrieve/obtain the cell lines or tissue sample from fresh or preserved sources and prepare a culture
  2. Select specified culture media and add any necessary growth agents or nutrients
  3. Incubate cells/tissue in specified conditions
  4. Inoculate the media with the specified amount of sample
  5. Monitor growth of tissue and cell lines and products
  6. Detect contamination and troubleshoot materials, equipment and techniques
  7. Passage samples by subculturing to preserve or grow the line
  8. Harvest cells or cell products to optimise yields
Retrieve/obtain the cell lines or tissue sample from fresh or preserved sources and prepare a culture

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Select specified culture media and add any necessary growth agents or nutrients

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Incubate cells/tissue in specified conditions

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Inoculate the media with the specified amount of sample

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Monitor growth of tissue and cell lines and products

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Detect contamination and troubleshoot materials, equipment and techniques

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Passage samples by subculturing to preserve or grow the line

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Harvest cells or cell products to optimise yields

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Preserve cells and tissues

  1. Select the appropriate preservation method
  2. Preserve the cell lines or tissue in accordance with the method
  3. Check preserved cell lines regularly to ensure viability is maintained
Select the appropriate preservation method

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Preserve the cell lines or tissue in accordance with the method

Completed
Date:

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Check preserved cell lines regularly to ensure viability is maintained

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain records

  1. Maintain records of batches of media and test data
  2. Maintain records of active and stored tissue and cell lines
  3. Ensure records are retrievable, legible and accurate
  4. Ensure records conform to the information management, records, quality system and legal requirements
Maintain records of batches of media and test data

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain records of active and stored tissue and cell lines

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure records are retrievable, legible and accurate

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure records conform to the information management, records, quality system and legal requirements

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Assessed

Teacher: ___________________________________ Date: _________

Signature: ________________________________________________

Comments:

 

 

 

 

 

 

 

 

Instructions to Assessors

Evidence Guide

The Evidence Guide provides advice on assessment and must be read in conjunction with the performance criteria, required skills and knowledge, range statement and the Assessment Guidelines for the Training Package.

Overview of assessment

Critical aspects for assessment and evidence required to demonstrate competency in this unit

Assessors should ensure that candidates can:

work safely and satisfy all legal and regulatory requirements, including the use and care of safety cabinets

demonstrate chain of custody for all cells, cell lines and tissues

prepare, dilute and sterilise reagents and culture media that are fit for purpose

choose and justify appropriate media and substrate material based on cost, cleaning, sterilising and maintenance of cell growth

successfully passage cell cultures by subculturing

grow cell lines and tissue to specifications without contaminating the original sample and the environment

count cells, identify a wide range of cell types and contaminants and recognise normal and abnormal cells

monitor cell growth and recognise and troubleshoot problems, such as contamination

store cells so that they remain viable

maintain accurate, traceable records of cell lines and tissues and logs of procedures and work completed.

Context of and specific resources for assessment

This unit of competency is to be assessed in the workplace or simulated workplace environment.

This unit of competency may be assessed with:

MSL933001A Maintain the laboratory/field workplace fit for purpose.

Resources may include:

laboratory equipped with appropriate test equipment/instruments, standards and reagents

enterprise procedures and standard methods

relevant tissues and cell lines.

Method of assessment

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.

Where applicable, reasonable adjustment must be made to work environments and training situations to accommodate ethnicity, age, gender, demographics and disability.

Access must be provided to appropriate learning and/or assessment support when required.

The language, literacy and numeracy demands of assessment should not be greater than those required to undertake the unit of competency in a work like environment.

This competency in practice

Industry representatives have provided the case studies below to illustrate the practical application of this unit of competency and to show its relevance in a workplace setting.

Biotechnology

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.

Biomedical

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 37°C 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.

Required Skills and Knowledge

Required skills

Required skills include:

working safely and satisfying all legal and regulatory requirements

preparing, diluting and sterilising reagents and culture media

choosing media and substrate material based on cost, cleaning, sterilising and maintenance of cell growth

passaging cell cultures by subculturing

growing cell lines and tissue to specifications without contaminating the original sample and the environment

counting cells, identifying a wide range of cell types and contaminants and recognising normal and abnormal cells

monitoring cell growth and recognising and troubleshooting problems

storing cells so that they remain viable

demonstrating chain of custody for all cells, cell lines and tissues

maintaining accurate, traceable records of cell lines and tissues and logs of procedures and work completed

Required knowledge

Required knowledge includes:

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 and viability

cell biology (structure, physiology, function, physiological cell growth requirements, nutrient requirements, respiration, temperature and 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 substrates (e.g. solid, semi-solid, gel or sponge, glass, disposable plastics and three-dimensional matrices)

techniques for pre-treating substrates (e.g. feeder layers, chemical treatments, such as poly D-lysine, collagen, gelatine and fibronectin)

role of ingredients in media (e.g. salts, carbohydrates, amino acids, vitamins, hormones, growth factors, serum and 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

Range Statement

The range statement relates to the unit of competency as a whole. It allows for different work environments and situations that may affect performance. Bold italicised wording, if used in the performance criteria, is detailed below. Essential operating conditions that may be present with training and assessment (depending on the work situation, needs of the candidate, accessibility of the item, and local industry and regional contexts) may also be included.

Codes of practice

Where reference is made to industry codes of practice, and/or Australian/international standards, it is expected the latest version will be used

Standards, codes, procedures and/or enterprise requirements

Standards, codes, procedures and/or enterprise requirements may include:

Australian and international standards, such as:

AS 1678 Emergency procedure guide -Transport

AS 2252 Biological safety cabinets

AS ISO 17025-2005 General requirements for the competence of testing and calibration laboratories

AS/NZS 2243 Set:2006 Safety in laboratories set

AS/NZS 2982.1:1997 Laboratory design and construction - General requirements

AS/NZS 4187:2003 Cleaning, disinfecting and sterilising reusable medical and surgical instruments and equipment, and maintenance of associated environments in health care facilities

AS/NZS ISO 14000 Set:2005 Environmental management standards set

AS/NZS ISO 9000 Set:2008 Quality management systems set

Australian code of good manufacturing practice for medicinal products (GMP)

Australian Dangerous Goods Code

client and product specifications

enterprise procedures, standard operating procedures (SOPs) and quality assurance procedures

gene technology regulations

Guide to physical containment levels and facility types

HB 9-1994 Occupational personal protection

laboratory manuals

manufacturer's instructions or verbal direction from laboratory manager, supervisor or senior technician

material safety data sheets (MSDS)

National Code of Practice for the labelling of workplace substances [NOHSC:2012 (1994)]

occupational health and safety (OHS) national standards and codes of practice

operation and maintenance manuals for automated media preparation equipment

quality assurance procedures

principles of good laboratory practice (GLP)

production schedules and instructions

Therapeutic Goods Regulations 1009

verified test methods

Hazards

Hazards may include:

biohazards, such as infectious agents and oncogenic DNA

chemical and radiation hazards

allergenic factors

cryogenic liquids, such as nitrogen

heat from burners and molten agar

ultraviolet (UV) light

sharps

contaminated clothing

Hazard control measures

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, MSDS 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, gowns, body suits and 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

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 and carbon dioxide

binocular inverted microscope

centrifuges

cryogenic vessels and transfer equipment, and liquid nitrogen

Selection criteria for media, materials and equipment

Selection criteria for media, materials and equipment may include:

costs

ease of cleaning or sterilisation

maintenance of cell growth

Pre-use checks

Pre-use checks include:

performing routine maintenance

checks on raw materials and consumables, including use by date, possible contamination and storage conditions

Cells and tissues

Cells and tissues may include:

animal cell lines, such as hybridoma, liver, epidermal, lymphoblastic and fibroblastic

plant cell lines, such as tobacco, arabidopsis, soya bean, tomato, roses and meristomatic tissue

yeasts

fungi

sperm, ova and embryos

adherent and suspension cultures

Preparing a primary culture

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, micromanipulation, use of selective media, density gradient centrifugation, selective adhesion techniques and selective detachment

Monitoring growth of tissue and cell lines

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( e.g. by Giemsa)

karyotype analysis

Preservation of cell lines

Preservation of cell lines may include:

freezing

cryopreservation (dry ice and liquid nitrogen)

Records

Records may involve:

paper or laboratory information management systems (LIMS)

cataloguing of all cell lines

stock levels

viability test results

Occupational health and safety (OHS) and environmental management requirements

OHS and environmental management requirements:

all operations must comply with enterprise OHS and environmental management requirements, which may be imposed through state/territory or federal legislation - these requirements must not be compromised at any time

all operations assume the potentially hazardous nature of samples and require standard precautions to be applied

where relevant, users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council (NHMRC) and State and Territory Departments of Health