Michael Faraday ( ) British Prime Minister William Gladstone, observing Faraday performing a particularly unlikely experiment one day, pointedly asked him how useful such a 'discovery' could possibly be. "Why," Faraday smartly replied, "you will soon be able to tax it!"

33 Years...

Ovulation Induction Ovulation Induction CVS CVS ICSI ICSI Cryopreservation of gametes/embryos Cryopreservation of gametes/embryos Preimplantation Genetic Diagnosis Preimplantation Genetic Diagnosis Stem Cell Technologies Stem Cell Technologies

Embryonic Stem Cells Amniotic Stem Cells Placenta-derived Stem Cells Fetal Stem Cells Cord Blood Stem Cells Adult Stem Cells Embryonic Stem Cells Amniotic Stem Cells Placenta-derived Stem Cells Fetal Stem Cells Cord Blood Stem Cells Adult Stem Cells

Reproductive Organ Tumors Reproductive Organ Tumors Urinary Insufficency Urinary Insufficency Endometriosis Endometriosis Infertility Infertility Reproductive Organ Tumors Reproductive Organ Tumors Urinary Insufficency Urinary Insufficency Endometriosis Endometriosis Infertility Infertility Fertility Preservation Fertility Preservation Infertility Treatment Infertility Treatment Germline Gene Therapy Germline Gene Therapy Fertility Preservation Fertility Preservation Infertility Treatment Infertility Treatment Germline Gene Therapy Germline Gene Therapy

Gargett, 2007 “Endometrial regeneration is established with the help of the cells in basalis, not functionalis” ( ( Prianishnikov, 1978; Padykula et al., 1984; Padykula,1991)

In sex-mismatched transplantation patients chimerism was observed in several tissues including liver, intestines and endometrium.In sex-mismatched transplantation patients chimerism was observed in several tissues including liver, intestines and endometrium. Kleeberger et al., 2002; Okamoto et al. 2002; Taylor, 2004

Gargett and Masuda, 2010

Gargett and Guo, 2010

“Self-renewal” “Stem Cell” “Progenitor Cell” “Transiently Amplified Cells” “Differentiated Cells” Differentiation Pluripotency

Nagano. 2007

Stem Cells Germline Gene Therapy Cell/Tissue Freezing In vitro Gamete Production Oocyte Sperm In vitro Culture Chemotherapy/ Radiotherapy Cell/Tissue Freezing Transplantation In vivo gamete production Fertility Preservation Infertility Therapy Gene Therapy

Kubota and Brinster,2005; Geens et al., 2008

Gamete Stem Cells -In vitro Culture- In vitro growth of murine spermatogonial stem cells. Kubota et al.,2003;2004

GSC- In vitro Culture 2005

Hayashi et al. 2011

Propagation of human spermatogonial stem cells in vitro Sadri-Adrekani et al. 2009, 2011

In vitro production of fertile sperm from murine spermatogonial stem cell lines Sato et al “The system can induce spermatogenesis from SSCs that show spermatogenic failure due to a micro-environmental defect in their original testes. Thus, an in vitro system is established that can be used to correct or manipulate the micro- environmental conditions required for proper spermatogenesis from murine SSC lines.”

Nagano. 2007

Evans and Kaufman, 1981 (mESC) Thompson et al., 1998 (hESC)

Sperm Donation Oocyte 8-cell embryos Morula Arrested embryos Embryo biopsy for PGD Affected embryos Blastocysts Blastomeres (In vitro culture) ICM Human embryonic stem (hES) cells + In Vitro Fertilization For ART purposes In Vitro Fertilization For research purposes Morula Parthenogenetic activation of oocytes SCNT Somatic cells Coculture with genetically-labelled cells (Findikli and Vemuri. 2009)

Sperm cells from embryonic stem cells

Testicular cell conditioned medium supports differentiation of embryonic stem(ES) cells into ovarian structures containing oocytes Lacham-Kaplan et al Derivation of oocytes from mouse embryonic stem cells Hubner et al. 2003

Sperm cells from embryonic stem cells Nayernia et al ICSI was applied on 210 oocytes 65 oocytes were fertilized 7 transgenic live birth

Findikli et al. 2006

Sperm Donation Oocyte 8-cell embryos Morula Arrested embryos Embryo biopsy for PGD Affected embryos Blastocysts Blastomeres (In vitro culture) ICM Human embryonic stem (hES) cells + In Vitro Fertilization For ART purposes In Vitro Fertilization For research purposes Somatic cell Reprogramming IPs Morula Parthenogenetic activation of oocytes SCNT Somatic cells Coculture with genetically-labelled cells (Findikli and Vemuri. 2009)

Oocyte 8-Cell Embryo Blastocyst hES Cells Somatic Cell Reprogramming iPSCs Morula Parthenogenetic Activation SCNT Somatic cells Embryonic Germ Cells Genetic Modification Genetic Selection In Vitro Differentiation Oocyte Sperm Adult Stem Cells 1 23

“Can bone marrow or peripheral blood-derived stem cells be sources of oocytes?” Oocytes from Adult Stem Cells “Oocyte generation in adult mammalian ovaries by putative germ cells in bone marrow and peripheral blood” Johnson et al Mice were sterilized by chemotherapy. Oocyte production was observed after peripheral blood transfusion.

“Germline stem cells and folicular renewal in the postnatal mammalian ovary” Johnson et al “Oogenesis in cultures derived from adult human ovaries” Bukovsky et al "Ovulated oocytes in adult mice derived from non-circulating germ cells" Eggan et al Oocytes from Adult Stem Cells

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Erasure of genomic imprinting??? Cell-cycle control & Differentiation ??? Marques-Mari et al. 2009

Jules Verne ( ) !" “Anything one man can imagine, other men can make real”!"

Studies show that gamete production from stem cells is now possible to some extent. On the other hand there are still numerous unresolved or unknown issues regarding the molecular pathways leading to the optimal production, maturation and functionality of the produced gametes. Nearly all studies have been performed on animal models. Therefore, whether similar success can be obtained in humans is still not clear. We are still couple of steps far from seeing their potentials in a clinical setting.