the future ivf lab
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Future developments in the IVF lab
Giles Palmer Mitera IVF, Athens
Future developments in the IVF lab
Areas of interest• The laboratory• Embryo culture • Embryo selection
• Fertility preservation
Tomorrow• Biotech Arena• Integration • Automation
IVF labs generally resist change - maintain consistencyCost/benefit
Research grants
Future developments in the IVF lab
The story so far…..
• Change from apprentice system• Accreditation/documentation• Commercially available media and instruments• QMS/QC cornerstone of todays IVF labs• Purpose built labs ?
Future developments in the IVF lab
Regulations and the search llence• EUTCD 2004- specific quality
and safety requirements with a critical point in the doctrine being clean air
• Air quality and VOCs (Cohen 1997, Legro 2010)
• Clean room environment-”protect the manufactured product”
Future developments in the IVF lab
Regulations and the search flence• EUTCD 2004- specific quality
and safety requirements with a critical point in the doctrine being clean air
• Air quality and VOCs (Cohen 1997, Legro 2010)
• Clean room environment-”protect the manufactured product”
Future developments in the IVF lab
Laboratory equipment
Future developments in the IVF lab
Laboratory equipment
Future developments in the IVF lab
Laboratory equipment
Future developments in the IVF lab
Heated glass worktop, integrated RFID antenna, touch screen interface
Future developments in the IVF labAutomation of ICSI• Since 1992 : male infertility• Experience operator, highly skilled
• Auto injection – transgenics • Use of MEMS and micro-robotics,
Nano-newton force sensors, Servo optical control
• RICSI : 7200 per second with an accuracy of 0.240, Immobilization of sperm in 6-7 seconds (CASA adapted)
• Survival rate 90% (Sun 2011)• Human trials on going
Future developments in the IVF lab
Automation of ICSI• Since 1992 : male infertility• Experience operator, highly skilled
• Auto injection – transgenics • Use of MEMS and micro-robotics,
Nano-newton force sensors, Servo optical control
• RICSI : 7200 per second with an accuracy of 0.240, Immobilization of sperm in 6-7 seconds (CASA adapted)
• Survival rate 90% (Sun 2011)• Human trials on going
Future developments in the IVF labAutomation of ICSI• Since 1992 : male infertility• Experience operator, highly skilled
• Auto injection – transgenics • Use of MEMS and micro-robotics, Nano-
newton force sensors, Servo optical control
• RICSI : 7200 per second with an accuracy of 0.240, Immobilization of sperm in 6-7 seconds (CASA adapted)
• Survival rate 90% (Sun 2011)• Human trials on going
• Robot assisted oocyte retrieval!!
Future developments in the IVF lab
Further developments of in vitro culture
• 40 years on…Biggers, Brinster• Basic physiological principles of Leese, Quinn and Gardner• In vitro culture sub-optimum (Schieve 2002, Richter 2008)• Call for “new generation media” with bio active factors
GM-CSF
Future developments in the IVF lab
• GM-CSF in IVF medium improves success rates in Recurrent Implantation Failure patients(Spandorfer et al., Am J ReprodImmunol 2008)
• GM-CSF is reduced in recurrent miscarriage patients(Perricone et al Am J Reprod Immunol 2003)
• Follicular fluid GM-CSF is reduced in women experiencing unexplained infertility(Calogero et al., Cytokine 1998)
Positive effect on cell number, implantation, blastocyst formation, regulates apoptosisRobertson 2011, Sjoblom 1999
GM-CSFLIFPAFGHIGF-IIGF-IIEGFTGFβ
TNFαIFNγ
Future developments in the IVF lab
Many other compounds possible inclusion culture media? lipids/ prostoglandins
Future developments in the IVF labStatic culture methods• Polystyrene tubes and dishes• Accumulation of toxins, free radicles
Systems to mimic reproductive tract• Microfluidics- technology of handling small
volumes of liquids “Lab on a chip”
• Application in co-culture (Mizuno 2007)• Sperm sorting, fertilization (Suh 2006)• Cumulus removal (Zeringue 2004)• Dynamic micro-funnel device enhancing
mouse embryo development (Heo 2010)
• Possibility of real time measurements?
Future developments in the IVF lab
• Further developments of in vitro culture
Future developments in the IVF lab
• Further developments of in vitro culture
m
Future developments in the IVF lab
Selecting for success
• How to predict viability?• Cell morphology• Early cleavage, polar body
orientation and pronuclei morphology
Future developments in the IVF lab
Selecting for successInvasive Genetic analysis has evolved with IVF techniques. • Euploid selection (PGS) not fulfilled expectations• Global view- array CGH, SNP, New generation sequencing • RCT underway….promising (Yang 2011)
• Search for non invasive molecular techniques• DNA array –cumulus cells (Assou 2008), Follicular fluid (Hamel 2010), Proteomic
approach to sex selection (Picton 2010)
• Closest to implimentation –Systematic study of the unique chemical fingerprint that in vitro embryo leave behind
• Compulsory embryo biopsy & PGS????
Future developments in the IVF lab
Selecting for successInvasive Genetic analysis has evolved with IVF techniques. • Euploid selection (PGS) not fulfilled expectations• Global view- array CGH, SNP, New generation sequencing • RCT underway….promising (Yang 2011)
• Search for non invasive molecular techniques• DNA array –cumulus cells (Assou 2008), Follicular fluid (Hamel 2010),
Proteomic approach to sex selection (Picton 2010)
• Closest to implimentation –Systematic study of the unique chemical fingerprint that in vitro embryo leave behind
• Compulsory embryo biopsy & PGS????
Future developments in the IVF lab
Selecting for successInvasive Genetic analysis has evolved with IVF techniques. • Euploid selection (PGS) not fulfilled expectations• Global view- array CGH, SNP, New generation sequencing • RCT underway….promising (Yang 2011)
• Search for non invasive molecular techniques• DNA array –cumulus cells (Assou 2008), Follicular fluid (Hamel 2010), Proteomic
approach to sex selection (Picton 2010)
• Closest to implimentation –Systematic study of the unique chemical fingerprint that in vitro embryo leave behind
• Compulsory embryo biopsy & PGS????
Future developments in the IVF lab
Selecting for successInvasive Genetic analysis has evolved with IVF techniques. • Euploid selection (PGS) not fulfilled expectations• Global view- array CGH, SNP, New generation sequencing • RCT underway….promising (Yang 2011)
• Search for non invasive molecular techniques• DNA array –cumulus cells (Assou 2008), Follicular fluid (Hamel 2010), Proteomic
approach to sex selection (Picton 2010)
• Closest to implimentation –Systematic study of the unique chemical fingerprint that in vitro embryo leave behind
• Compulsory embryo biopsy & PGS????
Future developments in the IVF lab
Selecting for successInvasive Genetic analysis has evolved with IVF techniques. • Euploid selection (PGS) not fulfilled expectations• Global view- array CGH, SNP, New generation sequencing • RCT underway….promising (Yang 2011)
Search for non invasive molecular techniques• DNA array –cumulus cells (Assou 2008), Follicular fluid (Hamel 2010), Proteomic
approach to sex selection (Picton 2010)
• Closest to implementation –Systematic study of the unique chemical fingerprint that in vitro embryo leave behind in culture media
• Compulsory embryo biopsy & PGS????
Future developments in the IVF lab
• Leese & Conaghan- late 1980’s- pyruvate uptake and embryo viability
• Bio-spectroscopy-applied to spent media (NMR/MS & HPLC) Houghton 2002, Brison 2004, Katz-Jaffe 2006
Future developments in the IVF lab
• Sakkas- biochemical factors representative of embryos that give pregnancy or no pregnancy
• 2009: Reduced complexity-Raman/NIR spectroscopy - rapid metabolic profile=Viability scores
• Automative system of embryo metabolisms – problems in its development
• Pipeline “Embryosure “- Amino acid profiling (Origio/Leese)
• Incorporation in microfluidic devices? (Swain 2009)
Future developments in the IVF lab
Time lapse evaluation
Morphological selection criteria in literature:
Cell divisionCell symmetry SynchronicityMulti-nucleationFragmentation
Abnormal cell division
Future developments in the IVF lab
Time lapse evaluation
Morphological selection criteria in literature:
Cell divisionCell symmetry SynchronicityMulti-nucleationFragmentation
Abnormal cell division
m
Future developments in the IVF lab
Time lapse evaluation• Use of morphokinetics as a predictor of embryo implantation (Meseguer 2011)
• Search for algorithm for implantation potential• Morphokinetics link to aneuploidy detection (Davies 2012)• Classification system based on time parameters relates to selection of euploidy embryo ( Basile
2013)• Applications: Oxygen consumption (lopes 2005), cell membrane tracking ( Wong 2010)
t2 t3 t4 t5
cc2 cc3s2
Cc: cell cyclecc2 = t3-t2cc3 = t5-t4s2 = t4-t3
cc2=11,8h s2 = <0.76h
t2=25,6h (24,3-25,8h)t3=37,4h (35,4-37,8h)t4=38h (36,4-38,9h)t5=52,3h (48,8-56-6,h)
Future developments in the IVF lab
Time lapse evaluation• Use of morphokinetics as a predictor of embryo implantation (Meseguer 2011)
• Search for algorithm for implantation potential• Morphokinetics link to aneuploidy detection (Davies 2012)• Classification system based on time parameters relates to selection of euploidy embryo ( Basile
2013)• Applications: Oxygen consumption (lopes 2005), cell membrane tracking ( Wong 2010)
t2 t3 t4 t5
cc2 cc3s2
Cc: cell cyclecc2 = t3-t2cc3 = t5-t4s2 = t4-t3
cc2=11,8h s2 = <0.76h
t2=25,6h (24,3-25,8h)t3=37,4h (35,4-37,8h)t4=38h (36,4-38,9h)t5=52,3h (48,8-56-6,h)
“Lab on a chip”
Sperm sorting
Imaging
Co-culture
Oxygen consumption
Biomarker analysis
Future developments in the IVF lab
Extra slides
The IVF lab of the futureAutomation in the ART. (mes)
• Mostly manual/sperm selection/oocyte selection/denudation/ICSI/monitoring/vitrification
• Emerging technology-alternative in all fields of medicine/ telepresence/education
• Da Vinci/AESOP/Zeus
• Robotic assisted follicle aspiration!
The IVF lab of the future
Fertility Preservation• Emerging medical discipline• Ovarian stimulation/Oocyte freezing may not be always appropriate
Options for cancer patients• IVM of primordial follicles• Xenographing• Cryopreservation/transplantation of ovarian tissue (Donnez 2004)Ovarian tissue culture, IVG, folliculogenesis• Live births with pre-antral primordial follicles in mice (Eppig 1996)• Advances in tissue/biomaterial engineering• 2D vs 3D models (alginate, poly ethylene glycol PEG)• Promoted secondary follicles to antral and MII in primates (Xu 2011)• Artificial ovary-thecal and granulosa cell self assemble (Krotz 2010)Folliculagenesis• Complex process-endocrine & paracrine interactions
The IVF lab of the future
Fertility Preservation
Perfusion system –pulsatile gonadotrophin administration mimic pituitary-enhances follicular developmentWinkler 2009,2013
The IVF lab of the future
Moore’s law• Intel based business
strategy• Exponential growth every 2
years• Generally difficult in
medicine• ART under utilized
• IVF labs generally resist change - maintain consistency
• Sakkas- biochemical factors representative of embryos that give pregnancy or no pregnancy
• Bio-spectroscopy- metabolic profile/ with bio-informatic analysis=Viability scores
• 2009-Automative system of embryo metabolisms – problems in its development
• Pipeline “Embryosure “- Amino acid profiling (Origio/Leese)
•Sakkas. Fertil Steril 90.6.(2008 )
The IVF lab of the future
The IVF lab of the future
The “omic” approach• Genomics, transciptomics, proteomics etc• Genetic analysis has evolved with IVF techniques• Euploid selection (PGS) not fulfilled expectations• Fgf• Bvv• Vcvc• Vcv• Cxcc• N
The IVF lab of the future
The “omic” approach• Genomics, transciptomics, proteomics etc• Genetic analysis has evolved with IVF techniques• Euploid selection (PGS) not fulfilled expectations• Fgf• Bvv• Vcvc• Vcv• cxcc
The IVF lab of the future
The “omic” approach• Genomics, transciptomics, proteomics etc• Genetic analysis has evolved with IVF techniques• Euploid selection (PGS) not fulfilled expectations• Fgf• Bvv• Vcvc• Vcv• cxcc
The IVF lab of the future
Laboratory equipment• Witness system, RFID• Microscopic silicon based barcoding (Novo 2010)
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