Transcript View Presentation - the lifeglobal® group

A Unified Approach to
Human Embryo Culture
Don Rieger PhD and
Klaus Wiemer Ph.D.
Life Global, Northwest Center for
Reproductive
Sciences
[email protected]
A Single Medium
vs.
Sequential Media
from Zygote to Blastocyst
A Single Medium vs. Sequential
Media for Embryo Culture
“… in order to optimize mammalian embryo
development in culture, sequential media are
required, each designed to meet the changing
requirements of the developing embryo.”
(Gardner and Lane, In Principles of Cloning, 187-213, 2002)
“The popularity of this conviction is due to its
intuitive appeal, but critical examination shows that
it is not supported by direct experimental evidence”
(Biggers et al., Hum Reprod 20, 3376-84, 2005)
 A single medium can be used
for culture from the zygote to
the blastocyst stage
 Changing the medium is a
stress on the embryo
The Development of global®
Medium
Year
Investigators
SOM
1991 Lawitts & Biggers
KSOM
1994 Erbach et al.
KSOMaa
2000 Biggers et al.
global® (Ala-Gln) 2002 Wiemer et al.
global® (Gly-Gln) 2006 (Biggers et al., 2004)
global® Formulation
Salts
NaCl
KCl
CaCl2
KH2PO4
MgSO4
Buffer
Energy Substrates
Non-Essential AA's
Glutamine Dipeptide
Essential AA's
Chelator
Indicator
Antibiotic
NaHCO3
Glucose
NaLactate
NaPyruvate
10 NEAA's
Gly-Gln
9 EAA's
EDTA
Phenol Red
Gentamicin
The
®
global
Family
of Media
The global® Family of Media
Objective
To maintain, as closely as possible, the
same medium throughout all stages of
gamete and embryo handling and culture
In order to
Minimize the stress of changing from one
medium to another
1.
®
global
for Fertilization
Energy Metabolism
of the COC and Sperm
global® for Fertilization Formulation
Components
Salts
Buffer
Energy Substrates
Non-Essential AA's
Glutamine Dipeptide
Essential AA's
Chelator
Indicator
Antibiotic
NaCl
KCl
CaCl2
global® for
Fertilization
NaCl
KCl
CaCl2
KH2PO4
KH2PO4
MgSO4
MgSO4
NaHCO3
Glucose
NaLactate
NaPyruvate
10 NEAA's
Gly-Gln
9 EAA's
EDTA
Phenol Red
Gentamicin
NaHCO3
Glucose (High)
NaLactate
NaPyruvate
10 NEAA's
Gly-Gln
9 EAA's
EDTA
Phenol Red
Gentamicin
global®
2. global® w/HEPES
Acid-Base Buffer Systems
global® w/HEPES Formulation
Components
Salts
Buffer
Energy Substrates
Non-Essential AA's
Glutamine Dipeptide
Essential AA's
Chelator
Indicator
Antibiotic
NaCl
KCl
CaCl2
global®
w/HEPES
NaCl
KCl
CaCl2
KH2PO4
KH2PO4
MgSO4
MgSO4
NaHCO3
Glucose
NaLactate
NaPyruvate
10 NEAA's
Gly-Gln
9 EAA's
EDTA
Phenol Red
Gentamicin
HEPES
Glucose
NaLactate
NaPyruvate
10 NEAA's
Gly-Gln
9 EAA's
EDTA
Phenol Red
Gentamicin
global®
3. PGD Biopsy Medium
The PGD Biopsy Procedure
The Effect of Biopsy Medium
on Pregnancy Rate
Preg. Rate (% of retrievals)
P < 0.05
50
40
30
20
10
0
Simple
No Biopsy
Biop.Medium
Time Period 1
No Biopsy
Supplemented
Biop. Medium
Time Period 2
(Hill & Li, The Clinical Embryologist 7, 11-12, 2004)
PGD Biopsy Medium Formulation
Components
global®
PGD Biopsy
NaCl
KCl
CaCl2
KH2PO4
MgSO4
NaCl
KCl
Non-Essential Amino Acids
NaHCO3
Glucose
NaLactate
NaPyruvate
10 NEAA's
HEPES
Glucose
NaLactate
NaPyruvate
10 NEAA's
Glutamine Dipeptide
Glycyl-glutamine
Glycyl-glutamine
Essential Amino Acids
9 EAA's
9 EAA's
Chelator
EDTA
EDTA
Indicator
Phenol Red
Phenol Red
Antibiotic
Gentamicin
Gentamicin
Salts
Buffer
Energy Substrates
Osmolyte
Protein
KH2PO4
Sucrose
HSA
The Unified Approach
to Culture
ART Procedures – Day 0
Classical IVF
ICSI
global® for Fertilization
global® w/HEPES
Culture
global®
Day 1
Day 1
ART Procedures – Day 1
Classical IVF
Post-ICSI Culture
global® for Fertilization
global®
Culture
fresh droplets of
global®
Day 3
ART Procedures – Day 3
Transfer
global®
Culture
global®
PGD Biopsy
PGD Biopsy Medium
Cryopreservation
Assisted Hatching
Culture
fresh droplets of
global®
Day 5
global® w/HEPES
ART Procedures – Day 5 and 6
Culture
Transfer
global®
Cryopreservation
global®
Culture
fresh droplets of
global®
Transfer
global®
Day 6
Cryopreservation
Thank You