GLP.pdf ART lab fertility Manipal academy of Higher education

ART

1. GoodLaboratoryPracticein ART Laboratory
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2. ARTsuccessrate
Failure
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Whatwentwrong?

3. ART
Stimulation
protocol
Gamete/
embryo
quality
Endometrial
receptivity
Laboratory
conditions
Culture
conditions
Skill
s
3
ART
Success

4. Goodlaboratorypractice
Plan Perform Monitor Record Analyze Report
•To ensure quality, reliability & integrity of the procedures conducted
•To improve the protection of human health and environment
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5. Components
ofGLP
Location
Staffing
work
culture
Personal
attitude
Instruments
Quality
control
Laboratory
ethics
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6. Laboratorystaffing
ClinicianEmbryologistNursing Technician
ReceptionistCounselorHousekeeping
DirectorQualifiedstaff
Skilledprofessionals
Adequatenumber
12
150cycles –2 Qualified Clinical Embryologists
supervisor

7. Laboratory director
•Higher academic degree-MD, MSc, PhD
•Minimum of 6 years of documented human embryology experience.
Responsibilities
•Selection and implementation of the most adequate materials and procedures
•Safe and appropriate laboratory facilities and equipment according to regulations.
•Implementation of a quality management system (QMS), laboratory risk management, and prevention policy.
•Sufficient laboratory staff members with the appropriate skills.
•A comprehensive orientation and introduction programmefor all new staff members.
•Management of laboratory staff training and continual scientific and biomedical education.
•Implementation and review of key performance indicators (KPIs)
•Approval of research projects by competent authorities.

8. Laboratory supervisors
Responsibilities
•Efficient organisationof daily work of their areas of responsibility
•Effective communication with laboratory staff and clinical colleagues.
•Continuous improvement where possible.
•Structured training of staff members and students.
•BSc in biomedical sciences
•3 years of documented human embryology experience

9. Clinical Embryologists•BSc in biomedical science
Responsibilities:
•Execution of standard operating procedures (SOPs).
•Participation in daily practice, communication, and organization.
•Contribution to laboratory clinical decisions.
•Training of staff members and students.

10. Timesense
Dedication/
Sincerity
Knowledge
Ethical
practice
Common
sense
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Personalattitude

11. Laboratorylocation
Awayfrompollutedplace
Canteen
Histologylab
Heavyconstructionsite
Commoncorridor
Goodaccesstothepatients
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12. Selectingthe CorrectFloorforIVFLab
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13. Laboratorydesign
Non
sterile
zone
Semi
sterile
zone
Sterile
zone
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14. Floor
Smoothsurface
Lowoffgassingmaterials
Easytowash
Non-reflective
Singlesheet
Nogaps
Curvededges
Polyvinylsheets
X
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√

15. Wall
Smoothsurface
Lowoffgassingmaterials
Easytowash
Non-reflective
Single sheet
Nogaps
Polyvinylsheets
Leadfreeepoxybasedpaints
Furniture-stainlesssteel
Window/door-aluminium
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16. Roof
Smoothsurface
Low offgassing materials
Easyto wash
Non-reflective
Single sheet
Nogaps
Polyvinylsheets
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17. Workculture
•Growinglongnails,nailpolish
•Usingdeodorant/perfumes/cosmetics/Hairstylinginsidelab
•Eating/drinkinginsidelab / Smoking/ alcoholconsumption
•Excessive talking/scratchingbodyparts
•Coughing/sneezing
X
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18. LaboratoryClothing
•Useseparatefootwear
•Wearaprotectiveclothing
•Exclusiveforlaboratory
•Shouldbecleanandneat
•Usenon-toxicgloves
Skullcap
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Mask
Laboratorycloth
Laboratorycloth
Footwear

19. Handwash
•Thoroughwashingwithclean water
•Milddetergents
•Noalcohol/strongsmellingdetergents
Removefinger ring, wristwatch,bangles
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20. •Dustparticles
•Pestsandinsects
•Airpollutants
•Fumesand badodors
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Protectsfrom
Air curtain

21. Independent airconditioner
Minimumhumantraffic
Avoid unnecessary movement in embryology
lab
Frequent opening and closing of embryology
complexdoors
Positive pressurelab
CODAtower
InternalAirquality
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22. Lightexposure
Nodirectexposuretovisible light
Light of blue wavelength –toxic
Microscopic light-blue filter
Dimmersin laboratory
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23. Temperature
Oocytes and embryos-highly sensitive
tofluctuatingtemperature
Disintegrationofspindlefibers
Maintain37°C-workbenchinlaminar
flow,microscopic stage,incubator
X √
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24. Instruments
Purchasing
Bestquality
Installing
Manufacturers
Using
Testperiod
Read
instruction
manual
Follow
manufacturers
instruction
Continuous
use
Maintaining
Cleaningafter
completionof
work
Regular
servicing
Neverfixthe
problem
yourself
RegularQC
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An automatic emergency backup power system must be in place
for all critical equipment.

25. Biologicalsafetycabinet
•Allowthecabinettooperate
unobstructedforatleast 30 min
•Surfacedecontamination-UV,alcohol
•Keepallmaterialsatleast fourinches
insidethe workbench 3-4inches
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26. Biologicalsafetycabinet
Comfortableseating
Chairwithwheels
Placingonlyhandsinsidetheflow
Keeping onlythe necessary
materialsinsidehood
Placing large objects close together
Talking
Peepinginsidethehood
Lettinghairloose√
X 23

27. Incubator
•Regularchecking of
•Waterlevel
•Microbialgrowth
•CO
2andtemperature
•Arranging the culturedishesneatly
•Putonlyyourhandinside
•Usingseparateincubatorsforsemen
•processing&embryoculture
•Cylinders should be kept outside the lab.
•High-purity gas and inline HEPA and VOC
filters are highly recommended.
•Frequentopeningandclosureofdoor
•Talking whenthedoorisopen
•Peepingintoincubator
•Spillingmediaorbiologicalfluid
Do’s
Don’ts
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28. Centrifuge
•Sturdycentrifuge
•Placingonsturdyplatform
•Balancingthetubeholders
•Using roundbottom tubes
•Using therightspeed
•Regularcleaning
•Keepingoverfilledtubes
•Openingthelidwhilecentrifugeisrunning
•Usingfragiletubes
•Runningwithoutbalance
•Applyingbrake
Fixedanglerotor
Swingingbucketrotor
Pellet
Pellet
Do’s
Don’ts
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29. Increasingefficiencyofequipments
Efficiency
testing
Specificbioassay
SatisfactoryPoor
Identify&correct
theproblem
Comparewithclinicalresult
Follow
GLP
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30. Culturemedium
Oocytesandembryos-highly sensitivetopH
andosmoticchanges
Checktheexpiry date
Storeat 4°C,make small aliquots
Keepthebottleupright
Acidic Alkaline
X
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√

31. Culturemedium
•Different approaches or culture systems can be used in order to optimize embryo development.
•Sequential or single-step media.
•The type and number of incubators should be appropriate to the workload.
•Oil overlay minimizes changes to temperature, Ph, and osmolality.
•For traceability purposes, single embryo culture is advisable.
•For blastocyst culture, a low oxygen concentration should be used

32. Microdropletculture
Paraffinoil
Medium
Oil
X
X
√
Prepare the dish at room
temperature
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33. Humidity
Ambientair-40to60%;Incubator>95%
Avoidsfungalgrowth
Optimum condition for equipment
performance
Comfortableworkenvironment
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34. Consumables
Use tissueculturegrade items
CE certified
Shouldbesterileanddisposable
Endotoxintested
Donotstorethe itemsforlong time
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35. Handlingcryogenicmaterial
Spectacles/eyeshield
Laboratorycloth
Cryogloves
•Causesoxygendeficient environment
•Workin well ventilatedroom
•Exclusiveuse forcryopreservation
•Protection
•Upon exposure-washwithwarm
water,exerciseto theexposedpart
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36. Labelingsystem
Permanentmarker
Printedlabels
Electronicbarcodingsystem
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37. ErrorfreeLab-Manualdoublewitnessingsystem
•Twopeoplewitnessinga
procedure
•Less chanceofmistakes
1.Eggcollection
2.Semencollection
3.Spermpreparation
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4.Oocytedenudation
5.Insemination
6.Fertilizationcheck
7.Embryotransfer
•Documentationofeachstep
8.Cryopreservation
9.Thawing

38. Laboratorycleaning
•Regularcleaning-Steriledeionizedwater
•Bodyfluid spillage-5.25%sodium hypochlorite
•Disinfection
•FumigationX
•Alcohol
•UVirradiation
•Testedcommercialbrands
•Floorcleaning
•Wetmop-
•SweepingX
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39. Instruments
Culture
medium
Consumables
Environment
Protocols
Personnel
Activities
requiringQC
Qualitycontrol
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•Partoftheprofession
•Prepareaschedule
•Involveentire team
•Documentthefindings
•Regularauditing
•InternalandexternalQC

40. GLP-summary
Understandthephysiologyofgametesandembryos
Punctuality,sincerity,dedication
Makechecklist
Regularmonitoring–instrument,skill,consumables,media
QC&QA
Do not useanyshortcuts
Aseptic handling&minimummanipulation
Minimumdisturbancestotheoocytesorembryosinculture
Practiceethics
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