Prof. Semra Kahraman M.D.Bio.Çağrı Beyazyürek, Zafer Candan, Sevil

Ünal, Semra Mılık, Assoc. Prof. Semih Özkan M.D.

Istanbul Memorial Hospital, ART and Reproductive Genetics Center

Istanbul, Turkey

Prof. Semra Kahraman M.D.Bio.Çağrı Beyazyürek, Zafer Candan, Sevil

Ünal, Semra Mılık, Assoc. Prof. Semih Özkan M.D.

Istanbul Memorial Hospital, ART and Reproductive Genetics Center

Istanbul, Turkey

CURRENT APPROACHES IN SEVERE MALE INFERTILITYCURRENT APPROACHES IN SEVERE MALE INFERTILITY

SEVERE MALE INFERTILITYSEVERE MALE INFERTILITY

Genetic factors

Preimplantation genetic diagnosis

Surgical sperm recovery techniques

Sperm DNA fragmentation

Derivation of gamete cells from embryonic stem cells

Development of artificial gametes

Genetic factors

Preimplantation genetic diagnosis

Surgical sperm recovery techniques

Sperm DNA fragmentation

Derivation of gamete cells from embryonic stem cells

Development of artificial gametes

Male Infertility and GeneticsMale Infertility and GeneticsMale Infertility and GeneticsMale Infertility and Genetics

Structural Chromosomal AbnormalitiesStructural Chromosomal Abnormalities

Translocations (Robertsonian, Reciprocal, Cryptic)Translocations (Robertsonian, Reciprocal, Cryptic)

Duplication, inversion, insertionDuplication, inversion, insertion

Numerical Chromosomal AbnormalitiesNumerical Chromosomal Abnormalities

Gain or loss of entire chromosomesGain or loss of entire chromosomes

Micro or macrodeletions on Y chromosomeMicro or macrodeletions on Y chromosome

Gene defectsGene defects

Structural Chromosomal AbnormalitiesStructural Chromosomal Abnormalities

Translocations (Robertsonian, Reciprocal, Cryptic)Translocations (Robertsonian, Reciprocal, Cryptic)

Duplication, inversion, insertionDuplication, inversion, insertion

Numerical Chromosomal AbnormalitiesNumerical Chromosomal Abnormalities

Gain or loss of entire chromosomesGain or loss of entire chromosomes

Micro or macrodeletions on Y chromosomeMicro or macrodeletions on Y chromosome

Gene defectsGene defects

GENETIC FACTORS İMH ART and Reproductive Genetics Center

GENETIC FACTORS İMH ART and Reproductive Genetics Center

Karyotype analysis of 1935 infertile men with severe oligozoospermia or azoospermia

1214 cases: Non-obstructive azoospermia (NOA)

721 cases: Severe oligoasthenoteratozoospermia (OAT) (total sperm concentration in the whole ejaculate< below 5 million).

(1364 cases: Y-microdeletion analysis)

Karyotype analysis of 1935 infertile men with severe oligozoospermia or azoospermia

1214 cases: Non-obstructive azoospermia (NOA)

721 cases: Severe oligoasthenoteratozoospermia (OAT) (total sperm concentration in the whole ejaculate< below 5 million).

(1364 cases: Y-microdeletion analysis)

RESULT RESULT In cases with severe male factor infertility the incidence of having an abnormality in at least one test (karyotype

analysis or Y-microdeletions) is 16,6% in our study.

In cases with severe male factor infertility the incidence of having an abnormality in at least one test (karyotype

analysis or Y-microdeletions) is 16,6% in our study.

Distribution of normal and abnormal karyotypes in infertile men

İMH ART and Reproductive Genetics Center

Distribution of normal and abnormal karyotypes in infertile men

İMH ART and Reproductive Genetics Center

KARYOTYPENOA

%OAT

%TOTAL

%n=1214 (%62) n=721(37.2) n=1935

Normal 973 80.15 645 89.46 1618 83.6

Klinefelter’s 133 10.95 5 0.69 138 7.13

Mosaic Klinefelter 10 0.82 6 0.83 16 0.83

Other sex chromosome mosaicism (45,X/46,XY)

13 1.07 2 0.28 15 0.78

45,X male 45,X,tas(Y;2)(p11.3;qter) 46,XX males

45,X male (1) 0.90 0 0.00 11 0.57

46,XX males (10)

Other Sex Chromosomal Abnormalities10 0.82 1 0.14 11 0.57

(isoXq, idic Y and 47,XYY)

Reciprocal Translocation 12 0.98 13 1.8 25 1.29

Robertsonian Translocation 1 0.08 12 1.66 13 0.67

Inversions 4 0.33 1 0.14 5 0.26

Markers 1 0.08 1 0.14 2 0.10

Other Abnormalities 4 0.33 1 0.14 5 0.26

Total Abnormalities 199 16.4 42 5.83 241 12.45

Normal variable features/ Heterochromatin Polymorphisms

42 3.46 34 4.72 76 3.93

Klinefelter Syndrome was the most frequently detected abnormality (57.3% of detected Klinefelter Syndrome was the most frequently detected abnormality (57.3% of detected

abnormalities)abnormalities)

Chromosomal Variants Chromosomal Variants

Heterochromatin polymorphism is considered as a variant of a normal karyotype, but is more frequent in infertile men.

More attention must be directed to infertile men with heterochromatin polymorphism

Heterochromatin polymorphism is considered as a variant of a normal karyotype, but is more frequent in infertile men.

More attention must be directed to infertile men with heterochromatin polymorphism

Y chromosome microdeletion results in severe male infertility

Y chromosome microdeletion results in severe male infertility

AZFc deletion 82%

NOA (%)NOA (%)n=1041n=1041

OAT (%)OAT (%)n=323n=323

TOTAL (%)TOTAL (%)n=1364n=1364

NORMALNORMAL 942942 317317 12591259

DELETEDDELETED 99 (9.5)99 (9.5) 6 (1.85)6 (1.85) 105 (7.7)105 (7.7)

AZFaAZFa 44 00 4 (3.8)4 (3.8)

AZFbAZFb 22 22 4 (3.8)4 (3.8)

AZFcAZFc 4848 44 52 (49.5)52 (49.5)

AZFbcAZFbc 2828 00 28 (26.6)28 (26.6)

AZFabcAZFabc 1717 00 17 (16.3)17 (16.3)

Y-microdeletion rate in several studies

Y-microdeletion rate in several studies

Country Patient Number %

Our study 1364 7.7

India 83 9.6

Spain 50 16.0

Japon 63 15.8

USA, Australia 50 20.0

USA 108 7.0

France 53 9.4

Finland 201 9.0

China 101 11.0

Slovenia 226 4.4

Taiwan 94 11.7

New Zealand 65 7.7

Country Patient Number %

Our study 1364 7.7

India 83 9.6

Spain 50 16.0

Japon 63 15.8

USA, Australia 50 20.0

USA 108 7.0

France 53 9.4

Finland 201 9.0

China 101 11.0

Slovenia 226 4.4

Taiwan 94 11.7

New Zealand 65 7.7

Patients having both karyotype abnormality and Y-micro-deletion Patients having both karyotype

abnormality and Y-micro-deletion

Karyotype Abnormalities Karyotype Abnormalities Regions deleted on Y-chromosomeRegions deleted on Y-chromosome NN % %

47,XXY47,XXY Partial AZFa, AZFb and AZFcPartial AZFa, AZFb and AZFc 11

46,XX male SRY+ (4)46,XX male SRY+ (4)

Y completeY complete 7746,XX male SRY- (2)46,XX male SRY- (2)

45,X male SRY+ (1)45,X male SRY+ (1)

46,X,del(Y)(q11.2)46,X,del(Y)(q11.2)del complete b, c (2)del complete b, c (2)

33del complete b, del partial c (1)del complete b, del partial c (1)

idic Y(p) idic Y(p)

del complete b, c, sy 160 (1)del complete b, c, sy 160 (1)

33del Y complete (1)del Y complete (1)

del partial a, del complete b, c (1)del partial a, del complete b, c (1)

46,XY(92%)/45,X(8%)46,XY(92%)/45,X(8%) del complete b, cdel complete b, c

88

45,X(22%),46,XY(78%)45,X(22%),46,XY(78%) del complete b, cdel complete b, c

45,X(43%),46,XY(57%)45,X(43%),46,XY(57%) del complete b, cdel complete b, c

mos45,X[9]/46,XY[40]/47,XYY[1]mos45,X[9]/46,XY[40]/47,XYY[1] del complete b, c, sy 160del complete b, c, sy 160

mos45,X(50%)/46,Xi(Y)(p11.1)(50%)mos45,X(50%)/46,Xi(Y)(p11.1)(50%) del Y complete, sry+del Y complete, sry+

mos45,X(24%)/46,X,idic(Yp)(72%)mos45,X(24%)/46,X,idic(Yp)(72%) del complete b, c del complete b, c

mos45,X(52%)46,X,idicY(p)(48%)mos45,X(52%)46,X,idicY(p)(48%) del complete b, c, sy 160del complete b, c, sy 160

45,X(30%))/46,XidicYp(70%)45,X(30%))/46,XidicYp(70%) del complete b, c, sy 160del complete b, c, sy 160

TOTALTOTAL    2222 (9.1.%) (9.1.%)

Why Y-chromosome Micro-deletion analysis before TESE procedure?

Why Y-chromosome Micro-deletion analysis before TESE procedure?

The deletion types of AZF a,b and c loci on Yq11, are the potential

prognostic factors in patients planned to undergo TESE/mic-TESE

procedures.

cAZF c = Approximately 50% of the cases, mature spermatozoa

cAZF b = Nearly impossible to find mature spermatozoa

cAZF b+c and c AZF a+b+c = Total absence of testicular spermatozoa

The deletion types of AZF a,b and c loci on Yq11, are the potential

prognostic factors in patients planned to undergo TESE/mic-TESE

procedures.

cAZF c = Approximately 50% of the cases, mature spermatozoa

cAZF b = Nearly impossible to find mature spermatozoa

cAZF b+c and c AZF a+b+c = Total absence of testicular spermatozoa

Y-microdeletion and Mic-TESE Y-microdeletion and Mic-TESE

Retreival type Retreival type n=41n=41 Deletion Deletion Sperm Sperm

recoveryrecoveryResult Result

Mic-TESEMic-TESE 30 patients / 30 patients / 35 35 cyclescycles

partial a (2) partial a (2)

partial c (25) partial c (25)

complete bc (2) complete bc (2)

partial abc (1) partial abc (1)

14 patients/14 patients/

16 mic-TESE 16 mic-TESE

SRR:(48.4%) SRR:(48.4%)

9 pregnancies9 pregnancies

PR:(64.2/patient)PR:(64.2/patient)

1 unembryonic/1 unembryonic/

1 clinical abort1 clinical abort

8 babies8 babies

EjaculateEjaculate 44partial c (3)partial c (3)

partial b (1)partial b (1)++

2 pregnancy, 2 pregnancy,

1 ET cancellation1 ET cancellation

ELSIELSI 11 complete ccomplete c ++ No pregnancyNo pregnancy

TESATESA 11 Complete cComplete c ++ No pregnancy No pregnancy

Sperm retreival rates of TESE patients with same micro-

deletions

Sperm retreival rates of TESE patients with same micro-

deletions

partial c (n=15) partial c (n=15) (152,157,158,254,255)(152,157,158,254,255)

Same deleted regionsSame deleted regions

(variable phenotypic (variable phenotypic expression)expression)

Sperm found (n=6) Sperm found (n=6) SRR: 40%SRR: 40%

No sperm (n=9)No sperm (n=9)

SRR:60%SRR:60%

Conclusions Conclusions The high frequencies of cytogenetic abnormalities and Y micro deletions definitely suggest the need for genetic screening and counselling in severe male factor cases.

Karyotyping should be regarded as a mandatory part of the pre-treatment screening process for all men referred for ICSI.

Y-deletion analysis test is neccessary before deciding TESE procedure.

The high frequencies of cytogenetic abnormalities and Y micro deletions definitely suggest the need for genetic screening and counselling in severe male factor cases.

Karyotyping should be regarded as a mandatory part of the pre-treatment screening process for all men referred for ICSI.

Y-deletion analysis test is neccessary before deciding TESE procedure.

Preimplantation Genetic

Diagnosis for Male Infertility

(Aneuploidy)

Preimplantation Genetic

Diagnosis for Male Infertility

(Aneuploidy)

Golden Standarts Golden Standarts Biopsy: Single Blastomere

Fixation: Hypotonic+Fixative method

Hybridization: FISH at least 9 chromosomes: 13,15,16,17,18,21,22,X,Y

Analysis at least 2 rounds+recheck

Transfer on day 4

Biopsy: Single Blastomere

Fixation: Hypotonic+Fixative method

Hybridization: FISH at least 9 chromosomes: 13,15,16,17,18,21,22,X,Y

Analysis at least 2 rounds+recheck

Transfer on day 4

MFMFMFMF

AMAAMAAMAAMA

Indications and combined factors in 1000 PGD cycles

Indications and combined factors in 1000 PGD cycles

RPLRPLRPLRPL

RIFRIFRIFRIF

15.9%15.9%

13.2%13.2%

(43.5%)(43.5%)

%9.2%9.2

5.8%5.8%

10.2%10.2% 14%14%

9.3%9.3%

3.8%3.8% 12%12%

1.1%1.1% 5.2%5.2%

RPLRPL

RIFRIF

AMAAMA

29.6%

n=129

29.6%

n=129

27.2%27.2%

11.6%11.6%

8.5%8.5%

2.5%2.5% 20.6%20.6%

Male Factor and PGD (n=433)Male Factor and PGD (n=433)

Sperm SourceSperm Source

Source n

Ejaculate 308

(71.1%)

mic-TESE/TESA 125

(28.9%)

Total 433

Number of cyclesNumber of cycles

55

33

146

66

76

12

0

20

40

60

80

100

120

140

160

NOA VA SOAT OAT AT IT

*

34%34%34%34%

20.4%20.4%20.4%20.4%

PGD for Only Male Factor Infertility

PGD for Only Male Factor Infertility

PGDPGD Without PGDWithout PGD P-valueP-value

No. of cycles No. of cycles 129129 263263

Female age (years)Female age (years) 30.5 30.5 ±±33..55 2828..33±±33..99 nsns

Mean MII oocytes Mean MII oocytes 13.2 13.2 ±±55..11 1212..99±±55..55 nsns

Fertilization rate (%)Fertilization rate (%) 73.573.5 70.370.3 nsns

Mean embryos Mean embryos transferedtransfered 2.42.4 3.13.1 <0.05<0.05

Pregnancy rate (%)Pregnancy rate (%) 58%58% 48.7%48.7% nsns

Abortion rate (%)Abortion rate (%) 7%7% 19.7%19.7% <0.05<0.05

Implantation rate (%)Implantation rate (%) 28.6%28.6% 13.8%13.8% <0.05<0.05

Istanbul Memorial Hospital ART and Genetics CenterIstanbul Memorial Hospital ART and Genetics Center

Effect of additional factors on the outcome of PGD cycles for male infertility

60 5864

38

72

26

0

10

20

30

40

50

60

70

80

abnormal embryos clinical pregnancy rate

%

MF

MF+AMA

MF+AMA+RIF

Distributing of Chromosomal Abnormality

Distributing of Chromosomal Abnormality

n n %%

CyclesCycles 433433

Mean maternal age, (min-max)Mean maternal age, (min-max) 33.5 (20-47)33.5 (20-47)

Embryos diagnosedEmbryos diagnosed

NormalNormal 4040

AbnormalAbnormal 6060

AneuploidAneuploid 77.877.8

MonosomyMonosomy 34.634.6

TrisomyTrisomy 30.430.4

Complex AneuploidyComplex Aneuploidy 30.830.8

OthersOthers 4.24.2

HaploidyHaploidy/polyploidy/polyploidy 22.222.2

PGD RESULTS Ejaculated vs Testicular

Sperm(Maternal ages below 38)

PGD RESULTS Ejaculated vs Testicular

Sperm(Maternal ages below 38)

EjaculateEjaculate

(SOAT)(SOAT)Testicular Testicular

Sperm (NOA)Sperm (NOA)

Cycles initiated Cycles initiated 5050 4545

Embryos diagnosed as Embryos diagnosed as abnormal, (%)abnormal, (%) 5656 6262

Clinical pregnancy, %Clinical pregnancy, % 64*64* 44.844.8

Conclusion Conclusion The results of our study shows that the rate of aneuploidy is as high as 60% in patients with severe male factor infertility

Aneuploidy rate increases with the presence of other combined contributing factors such of other combined contributing factors such as AMA, RIF and RSA as AMA, RIF and RSA

PR dramatically decreases as more indications are combined with male infertility.

The results of our study shows that the rate of aneuploidy is as high as 60% in patients with severe male factor infertility

Aneuploidy rate increases with the presence of other combined contributing factors such of other combined contributing factors such as AMA, RIF and RSA as AMA, RIF and RSA

PR dramatically decreases as more indications are combined with male infertility.

Preimplantation Genetic Diagnosis

(PGD)

Preimplantation Genetic Diagnosis

(PGD)TranslocationsTranslocations

ProbesProbesProbesProbes

Locus Spesific (LSI) (200-500kb)

Centromeric (CEP) (alpha satellite p11-q11)

Telomeric (Tel) (60-170kb)

Whole Chromosome Painting Probes (WCP)

Locus Spesific (LSI) (200-500kb)

Centromeric (CEP) (alpha satellite p11-q11)

Telomeric (Tel) (60-170kb)

Whole Chromosome Painting Probes (WCP)

PGD-translocations n=104PGD-translocations n=104PGD-translocations n=104PGD-translocations n=104

PGD for Male translocation Carriersn=104

PGD for Male translocation Carriersn=104

RobertsonianRobertsonian ReciprocalReciprocal

PatientsPatients 2929 4848

CyclesCycles 3737 6767

Biopsied embryosBiopsied embryos 215215 418418

With conclusive With conclusive resultsresults

186186 366366

AbnormalAbnormal 104 (55.9%)104 (55.9%) 291 (79.5%)291 (79.5%)

NormalNormal 82 (44.1%)82 (44.1%) 75 (20.5%)75 (20.5%)

Mean maternal Mean maternal ageage

32.7 (23-45)32.7 (23-45) 32.5 (20-47)32.5 (20-47)

ET cyclesET cycles 31 (83.8%)31 (83.8%) 48 (%71.6)48 (%71.6)

PR/ET cyclesPR/ET cycles 10/31 (32.2%)10/31 (32.2%) 11/48 (22.9%)11/48 (22.9%)

Sperm FISH Aneuploidy screening for translocation cases. Is There Any Interchromosomal Effect? (n=5)

Sperm FISH Aneuploidy screening for translocation cases. Is There Any Interchromosomal Effect? (n=5)

Disomy ratesDisomy rates

Karyotype Karyotype 1313 1818 2121 XYXY

11 46,XY,rcpt(9;18)(p13.3;q21.3)46,XY,rcpt(9;18)(p13.3;q21.3) 00 (27.4)(27.4) 0.20.2 11

22 46,XY,rcpt(3;17)(p13;q23)46,XY,rcpt(3;17)(p13;q23) 0.60.6 00 00 11

33 46,XY,rcpt(11;15)(p12;p13)46,XY,rcpt(11;15)(p12;p13) 11 00 00 11

44 45,XY robt(13;14)(q10;q10)45,XY robt(13;14)(q10;q10) (9.2)(9.2) 0.10.1 44 0.20.2

55 45,XY robt(13;14)(q10;q10)45,XY robt(13;14)(q10;q10) (9.5)(9.5) 0.20.2 0.40.4 0.30.3

Conclusion Conclusion . PGD should be a viable alternative for translocation carriers to reduce miscarriages

Spermatozoa FISH testing may be used as an indicator of aneuploidy and segregation rate in gametes in translocation carriers and can give good approximation of success in a PGD cycle

Aneuploidy screening should be a part of genetic evaluation if female partner is >38 years

. PGD should be a viable alternative for translocation carriers to reduce miscarriages

Spermatozoa FISH testing may be used as an indicator of aneuploidy and segregation rate in gametes in translocation carriers and can give good approximation of success in a PGD cycle

Aneuploidy screening should be a part of genetic evaluation if female partner is >38 years

Micro-Dissection TESE Procedures in

Azoospermic Patients

Micro-Dissection TESE Procedures in

Azoospermic Patients

SPERM RECOVERY in NOA PATIENTS (n=1023)

İMH Andrology Unit

SPERM RECOVERY in NOA PATIENTS (n=1023)

İMH Andrology Unit

Micro-TESE: (NOA) 729

TESA 294

Sperm recovery rate in NOA Cases 375 / 729 = % 51.4

Micro-TESE: (NOA) 729

TESA 294

Sperm recovery rate in NOA Cases 375 / 729 = % 51.4

Results of Mic-TESEResults of Mic-TESE

Patients with first Mic-TESE trials :– No of patients : 591– Sperm recovery: 354– No sperm : 237 Sperm recovery rate: 354 / 591 = (%60)

Patients with previously conventional TESE trial with no sperm recovery :– No of patients: 60– Sperm recovery: 32– No sperm: 28

Sperm recovery rate: 32 / 60 = %53.3

Patients with first Mic-TESE trials :– No of patients : 591– Sperm recovery: 354– No sperm : 237 Sperm recovery rate: 354 / 591 = (%60)

Patients with previously conventional TESE trial with no sperm recovery :– No of patients: 60– Sperm recovery: 32– No sperm: 28

Sperm recovery rate: 32 / 60 = %53.3

Secondary MicroTESE success rates in patients

Secondary MicroTESE success rates in patients

Secondary MicroTESE n=242

Sperm recovered: 178

No Sperm : 64

Success rate :178/242=73.5%

Secondary MicroTESE n=242

Sperm recovered: 178

No Sperm : 64

Success rate :178/242=73.5%

TESE/mic-TESETESE/mic-TESE

TESE 36% vs Mic-TESE (Schlegel) 68% TESE 36% vs Mic-TESE (Schlegel) 68%

Klinefelter’s SyndromeKlinefelter’s Syndrome

No of cases: 65 Micro-TESE cases

Sperm found: 26

No sperm : 39

Success rate 26/65 = % 40

No of cases: 65 Micro-TESE cases

Sperm found: 26

No sperm : 39

Success rate 26/65 = % 40

Surgical sperm recovery rate according to hystopathologySurgical sperm recovery rate according to hystopathology

Sertoli cell only (Germ cell aplasia)

No of cases: 56

Sperm found: 20

No sperm : 36

Success rate: 20/56 = % 35

Sertoli cell only (Germ cell aplasia)

No of cases: 56

Sperm found: 20

No sperm : 36

Success rate: 20/56 = % 35

Surgical sperm recovery rate according to hystopathologySurgical sperm recovery rate according to hystopathology

Maturation arrest

Total number of cases : 37

Sperm found : 19

No sperm found : 18

Success rate : 19 / 37 = 51%

Maturation arrest

Total number of cases : 37

Sperm found : 19

No sperm found : 18

Success rate : 19 / 37 = 51%

ConclusionConclusionMic-TESE is one of the most recent and

important advance in surgical sperm

retrieval techniques. It’s success in

sperm retrieval and less complication

rates made this technique is the most

preferable procedure in sperm retrieval.

Mic-TESE is one of the most recent and

important advance in surgical sperm

retrieval techniques. It’s success in

sperm retrieval and less complication

rates made this technique is the most

preferable procedure in sperm retrieval.

ConclusionConclusionMic-TESE procedure has been you

used in our clinic since June 2002. mic-

TESE operation improved our sperm

retrieval rates and fulfilled some of our

patients hopes of having biologically

their own offsprings.

Mic-TESE procedure has been you

used in our clinic since June 2002. mic-

TESE operation improved our sperm

retrieval rates and fulfilled some of our

patients hopes of having biologically

their own offsprings.

Sperm DNA Fragmantation

• Meta-analysis:SCSA, performed in semen, cannot predict the outcome of ICSI

(Evenson D. 2006)

Sperm DNA fragmantation/TUNEL TEST

Sperm DNA fragmantation/TUNEL TEST

Terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling assay.

To detect the DNA damage, accounting for apoptotic sperms

At least 500 sperm are count for evaluation

The clinical value of TUNEL in predicting of IVF/ICSI outcomes in terms of fertilization rate and clinical outcome is not clear yet;

– < 20 % low degree of sperm DNA damage group

– ≥ 20 % high degree of sperm DNA damage group

Terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling assay.

To detect the DNA damage, accounting for apoptotic sperms

At least 500 sperm are count for evaluation

The clinical value of TUNEL in predicting of IVF/ICSI outcomes in terms of fertilization rate and clinical outcome is not clear yet;

– < 20 % low degree of sperm DNA damage group

– ≥ 20 % high degree of sperm DNA damage group

Representative Images of TUNEL Asssay

Representative Images of TUNEL Asssay

Green stained ones are apoptotic sperms, high DNA fragmentation

Green stained ones are apoptotic sperms, high DNA fragmentation

DNA FragmantationDNA Fragmantation >20%>20% <20%<20%

No of patientsNo of patients 1818 1919

Mean Male Age Mean Male Age 35.735.7 37.637.6

Mean Sperm Concentration (mil/ml)Mean Sperm Concentration (mil/ml) 20.620.6 33.933.9

Mean Total Sperm Motility (%)Mean Total Sperm Motility (%) 23.023.0 30.430.4

Mean Progressive Motile Sperm (%)Mean Progressive Motile Sperm (%) 3.33.3 5.25.2

Tunel TEST in RIF Cases with Low Sperm Motility

Tunel TEST in RIF Cases with Low Sperm Motility

Tunel TEST in RIF Cases with Low Sperm Motility

Tunel TEST in RIF Cases with Low Sperm Motility

DNA FragmentationDNA Fragmentation ≥ ≥ 20%20% < 20%< 20% pp

nn 1818 1919

Mean ♀ AgeMean ♀ Age 32.5 32.5 ±± 6.4 6.4 32.2 32.2 ±± 4.7 4.7 nsns

Mean Mean ♂♂ Age Age 35.7 35.7 ±± 6.6 6.6 37.3 37.3 ±± 5.1 5.1 nsns

Fertilization RateFertilization Rate 75.2 %75.2 % 83.1 %83.1 % nsns

Slow Growing Embryo on Day3Slow Growing Embryo on Day3 30.1%30.1% 32.6%32.6% nsns

Mean of ET NumberMean of ET Number 2.52.5 2.42.4 nsns

Implanatation RatesImplanatation Rates 20 %20 % 19.5 %19.5 % nsns

Clinıcal Pregnancy RatesClinıcal Pregnancy Rates 25 %25 % 30 %30 % nsns

210 mouse oocyctes (ICSI):

65 fertilization

7 transgenic births

210 mouse oocyctes (ICSI):

65 fertilization

7 transgenic births Nayernia et al.,2006

This study shows the higher plasticity potential of adult stem cells, like hES cells

In April 2007, Nayernia declared that his team obtained the obtained the bone marrow stem cells, called mesenchymal stem cells, bone marrow stem cells, called mesenchymal stem cells, from four adult men who were about to undergo bone from four adult men who were about to undergo bone marrow transplantsmarrow transplants

BMS cells are able to differentiate to early germ cells, primordial germ cells (PGCs) and even spermatogonial stem cells (SSC) and spermatogonia in vitro and in vivo.

This study shows the higher plasticity potential of adult stem cells, like hES cells

In April 2007, Nayernia declared that his team obtained the obtained the bone marrow stem cells, called mesenchymal stem cells, bone marrow stem cells, called mesenchymal stem cells, from four adult men who were about to undergo bone from four adult men who were about to undergo bone marrow transplantsmarrow transplants

BMS cells are able to differentiate to early germ cells, primordial germ cells (PGCs) and even spermatogonial stem cells (SSC) and spermatogonia in vitro and in vivo.

In-vitro generated artificial gametes: ultimate solutionIn-vitro generated artificial

gametes: ultimate solution

Patients with absent Patients with absent gametes or gonadsgametes or gonads

Somatic cell haploidization. Converting somatic cells from mitotic division to meiotic division directly.

De-differentiating somatic cells into embryonic stem cell and re-differantiating ES cells into gametes.

Extracting adult stem cell and re-differentiating them into gametes.

Patients with absent Patients with absent gametes or gonadsgametes or gonads

Somatic cell haploidization. Converting somatic cells from mitotic division to meiotic division directly.

De-differentiating somatic cells into embryonic stem cell and re-differantiating ES cells into gametes.

Extracting adult stem cell and re-differentiating them into gametes.

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ASSISTED REPRODUCTIVASSISTED REPRODUCTIVTECHNIQUESTECHNIQUESDIRECTOR:DIRECTOR:

Prof. SEMRA KAHRAMAN MD.Prof. SEMRA KAHRAMAN MD.

IVF CLINICIVF CLINICSEMRA KAHRAMAN MD.SEMRA KAHRAMAN MD.

GÜVENÇ KARLIKAYA MD. GÜVENÇ KARLIKAYA MD. HALE KARAGÖZOĞLU MD.HALE KARAGÖZOĞLU MD.

AYNUR ERŞAHİN MD.AYNUR ERŞAHİN MD.MÜSTECEP KAVRUT MD.MÜSTECEP KAVRUT MD.

MUSTAFA ACET MD.MUSTAFA ACET MD.NUR DOKUZEYLÜL MD.NUR DOKUZEYLÜL MD.

ŞEREF SARICA MD.ŞEREF SARICA MD.CRYO / EMBRYO / ANDROLOGYCRYO / EMBRYO / ANDROLOGY

CO CULTURE LABORATORYCO CULTURE LABORATORYSEVIL UNAL Bio. SEVIL UNAL Bio.

HAKAN YELKE Bio. HAKAN YELKE Bio. GÜNSELİ CENGİZ Bio. GÜNSELİ CENGİZ Bio. ZAFER ATAYURT Bio.ZAFER ATAYURT Bio.YEŞİM KUMTEPE Bio. YEŞİM KUMTEPE Bio.

SEMRA MILIK Bio. SEMRA MILIK Bio. ŞEBNEM ÜNVER Bio. ŞEBNEM ÜNVER Bio.

ÖZLEM YUVACAN Bio. ÖZLEM YUVACAN Bio. FERHAT CENGİZ Bio. FERHAT CENGİZ Bio.

SERKAN SELİMOĞLU Bio. SERKAN SELİMOĞLU Bio. ANDROLOGYANDROLOGY

Assoc. Prof. SEMİH ÖZKAN Assoc. Prof. SEMİH ÖZKAN MD.MD.

PERINATOLOGYPERINATOLOGYCİHANGİR YILANLIOĞLU MD.CİHANGİR YILANLIOĞLU MD.

ALTUĞ SEMİZ MD.ALTUĞ SEMİZ MD.

REPRODUCTIVE GENETICSREPRODUCTIVE GENETICSFRANCESCO FIORENTINOFRANCESCO FIORENTINOPhD. GÜLAY ÖZGÖN MD.PhD. GÜLAY ÖZGÖN MD.

MOLECULAR GENETICSMOLECULAR GENETICSBAHAR İSMAİLOĞLU Bio.BAHAR İSMAİLOĞLU Bio.

SELMA DEMİRSELMA DEMİR

FISHFISH ÇAĞRI OĞUR Bio.ÇAĞRI OĞUR Bio.

ÇİĞDEM ÇINAR Bio.ÇİĞDEM ÇINAR Bio.

CYTOGENETICSCYTOGENETICS ÖZLEM ÖNER Bio.ÖZLEM ÖNER Bio.ÇİLEM ASLAN Bio.ÇİLEM ASLAN Bio.

RESEARCH ANDRESEARCH ANDDEVELOPMENTDEVELOPMENT

N.ZAFER CANDAN Bio. N.ZAFER CANDAN Bio.

PUBLIC RELATIONSPUBLIC RELATIONSKÜBRA BURNAZKÜBRA BURNAZ

PATIENT RELATIONSPATIENT RELATIONSZEHRA ÖZKAL ZEHRA ÖZKAL

YASİN İZGİYASİN İZGİ

I.T. DEPARTMENTI.T. DEPARTMENTAYHAN EMİNOĞLUAYHAN EMİNOĞLU

SİBEL BEYAZAYSİBEL BEYAZAY

İLETİŞİMİLETİŞİMYELİZ SOYDANYELİZ SOYDANCEREN ERDEMCEREN ERDEM

NURSINGNURSINGYASEMİN GÜLERYASEMİN GÜLERSAİME TEPEBAŞSAİME TEPEBAŞSELVER ÇİÇEKSELVER ÇİÇEKCANAN YILMAZCANAN YILMAZ

HATİCE ALDEMİRHATİCE ALDEMİRDERYA SİVRİDERYA SİVRİ

SAFİYE SARIKOÇSAFİYE SARIKOÇHANDE MUTLUHANDE MUTLUSEMA KANATSEMA KANAT

GÜLAY ERYİĞİTGÜLAY ERYİĞİTSEYHAN GÜNDÜZSEYHAN GÜNDÜZ

INFORMATIONINFORMATIONDERYA ŞAHİNDERYA ŞAHİN

SEVİL TAULLAHSEVİL TAULLAHAYŞEGÜL BEZKAYŞEGÜL BEZKSEVDA UYANIKSEVDA UYANIK

SELEN EMRE TURANSELEN EMRE TURANNURAN SEYVANNURAN SEYVANBÜŞRA DURMAZBÜŞRA DURMAZGÜLŞEN TINKIRGÜLŞEN TINKIRAYŞEN KALAYCIAYŞEN KALAYCI

İLKAY ALİLKAY AL

ARCHIVEARCHIVEMUHAMMED KENARMUHAMMED KENAR

RAMAZAN ÇALIKRAMAZAN ÇALIK

FINANCEFINANCEAYŞEGÜL BEZKAYŞEGÜL BEZKBETÜL YAVAŞBETÜL YAVAŞ