QUALITY CONTROL IN THE IVF LAB. HOW TO DO IT Gloria Calderón PhD

Factors affecting the success of ART Diagnosis and therapeutic guidance. Ovarian stimulation Oocyte retrieval IVF procedures. Embryo transfer. Luteal phase treatment.

Why? Laboratory improvements affects directly in the ART programmes results. Programme results are mostly affected by laboratory variations. Rackowsky et al. 11th World Congress on IVF and Human Reproductive Genetics. Sydney, Australia 1999.

Laboratory: Key points Human resources Laboratory facilities Quality control

Human resources 6 embryologists 10 technicians 1035 cycles/year

ENOUGH TIME FOR EVERY CYCLE Optimal ratio of laboratory staff to ensure high attention during the procedures CONCENTRATION Each sample must be handled individually. RESPONSABILITY For the correct application of the protocols in the laboratory during ART procedures

Laboratory: Key points Human resources Laboratory facilities Quality control

Own oocytes/Laboratory % Pregnancy % Implantation EQUIPO IVI 2001

ICSI / Embryologist % 2PN % Pregnancy % Implantation EQUIPO IVI 2001

ICSI / Microscope % Pregnancy % Implantation EQUIPO IVI 2001

ICSI / Incubator % Pregnancy % Implantation EQUIPO IVI 2001

Technical facilities: equipments LAMINAR FLOW HOODS: Satisfactory number to avoid 2 procedures at the same time. MICROSCOPES: Satisfactory number to avoid procedure delays. INCUBATORS: 1 incubator/2 patients + working incubators.

>18 hours delay in 2PN examination …… affects the number of fertilized oocytes Nagy y col Hum Reprod 9: 1743-8, 1994 Suppinyopong y col J med Assoc Thai 83: 500-3, 2001 > 6 hours between denudation and ICSI or oocyte retrieval and ICSI Oocyte activation significantly increased Fertilization rates significantly decreased Albert y col C-59, SEF 2000

IVF Laboratory: equipments Annual maintenance contracts of equipments and filters Daily control of Tª, CO2 and pH Daily check of clinical results: early detection of lab problems

Technical facilities: characteristics Stable culture conditions Mineral oil Warmed surfaces Open incubators as less as possible -

Cohen et al

Laboratory: Key points Human resources Laboratory facilities Quality control

Quality control Daily control of temperatures, CO2 pH Embryotoxicity tests Accreditation of the IVF laboratory

Refrigerators Warmed plates Incubators Daily record of temperatures, CO2 and pH Eeejelemeheheheheh Jjflflflflflfl lhihhk Refrigerators Warmed plates Incubators

Application of embryotoxicity tests Contact materials Culture medium -1 or 2 cells mouse embryos -Sperm motility Fleetham y col Fertil Steril 59: 192-6, 1993 Fleeming y col Fertil Steril 47: 858-60, 1987 Bavister y Andrews J In Vitro Fertil Embryo Transf 5: 67-75, 1989 Claassen y col Hum Reprod 15: 1586-91, 2000

EUROPEAN COOPERATION FOR ACCREDITATION Accreditation of the IVF laboratory Legislation and recommendations to ensure quality performance of ART laboratory procedures (USA, UK, Sweden) ESHRE, ASRM, ALPHA EUROPE EUROPEAN COOPERATION FOR ACCREDITATION www.european-accreditation.org IVF LABORATORY´S TECHNICAL COMPETENCE

Audits Accreditation of the IVF laboratory IVF Laboratory Administrative questions Laboratory questions Clinic questions IVF Laboratory - Equipments calibration - Quality control documentation - Working manuals of the IVF methods - Documentation of results - Dated and signed reports Gianaroli y col Hum Reprod 15: 2241-46, 2000

Environmental factors   Environmental  factors Drugs manufacture Manufacture of accurate optical instrument What about us?

Determination of main pollutants compounds SO2 Petroleum hydrocarbons (C3-C9) Toluene Benzene Styrene and xylenes

Air quality influence 1994: Laboratory adjacent renovation and construction 1995: New system of air circulation 1993 n=71 1994 n=68 1995 n=60 1996 n=47 1997 n=29 p Pregnancies (%) 25 (35) 11 (16) 12 (20) 15 (32) 17 (59)* 0.003 Fertilization rate 74.1 60.2 62.4 70.7 69.2 0.032 % > 4 cell embryos 66# 61# 78# 77# 83# <0.001 * * • • Boone WR et al. Fertil Steril 1999; 71: 150-154

* * CODA No CODA 52 % 30 % % Pregnancy 40.5 % 37.1 % 1.4 % 14 % JF Mayer y col 1999, Norfolk USA 40.5 % 37.1 % 1.4 % 14 % * % Pregnancy % Miscarriage Racowsky y col 1999, Sydney Australia Torelló y col MEFS 2001 % Pregnancy % Miscarriage % Implantation 55 % 48.4 % 10.1% 9.4 % 34.3 % 27.7%

IVI-BARCELONA RESULTS (OCT2004-JAN2008) REGULAR RECIPIENTS PGD X AGE 35,07 40,8 38,2 Nb PROC 777 1216 248 Nb TRANSF. 649 1127 140 X EMB. TRANS 1,98 2,0 1,61 PREGNANCY % 54,1 61,3 42,8 IMPLANTATION % 37,5 43,7 35,2 MULTIPLE PREG. % 35,6 40,1 30,0