1. Establishment of autologous embryonic stem cells derived from preantral follicle culture and oocyte parthenogenesis 
Seung Tae Lee, Ph.D., Mun Hwan Choi, M.S., Eun Ju Lee, Ph.D., Seung Pyo Gong, B.Sc., Mi Jang, B.Sc., Sang Hyun Park, M.S., Hyang Jee, D.V.M., Dae Yong Kim, D.V.M., Ph.D., Jae Yong Han, Ph.D., Jeong Mook Lim, D.V.M., Ph.D. 
Fertility and Sterility 
Volume 90, Issue 5, Pages (November 2008)
DOI: /j.fertnstert
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Development of preantral follicles retrieved from the ovaries of F1 (C57BL/6 × DBA2) mice during in vitro culture. Mechanically retrieved secondary follicles were cultured in α-minimal essential medium-GlutaMAX medium supplemented with fetal bovine serum, insulin, transferrin, selenium, FSH, and antibiotics. (A) Follicular stage: the follicle remained spherical during culture, and a distinct basement membrane is visible (original magnification, ×600). (B) Diffuse stage: granulosa cells that enclose the oocyte have proliferated and grown out (original magnification, ×600). (C) Pseudoantral stage: the follicle has formed an antrum-like translucent structure as a result of the proliferation and differentiation of granulosa cells (original magnification, ×300). (D) Degenerative stage: the granulosa cells have degenerated after the oocyte spontaneously detached from the granulosa cell complex (original magnification, ×300). Bars = 50 μm in A, B; bars = 100 μm in C, D.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Characterization of follicle-derived, autologous embryonic stem (ES) cells (A) established by parthenogenetic activation and subsequent subculture of inner cell mass cell colonies in modified knockout Dulbecco's minimum essential medium supplemented with a 3:1 mixture of fetal bovine serum and knockout serum replacement. Follicle-derived mouse ES cells were characterized by using seven stem cell-specific markers: alkaline phosphatase (F) and anti-stage-specific embryonic antigen (SSEA)-1 (B), anti-SSEA-3 (C), anti-SSEA-4 (D), Oct-4 (E), anti-integrin α6 (G), and anti-integrin β1 (H) antibodies. The established ES cells stained positively with all of the specific markers, except anti-SSEA-3 and anti-SSEA-3, which share identity with mouse ES cells of other origins. Bars = 50 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Telomerase activities of the established follicle-derived, homozygous embryonic stem (ES) cells AS detected by the telomeric repeat amplification protocol assay. The ladder of telomerase products amplified by polymerase chain reaction was shown with six-base increments starting at 50 nucleotides, at the portion indicated (∗). The established ES cell line expresses high levels of telomerase activity. Lane 1, colony-forming cells of follicle-derived, homozygous ES cells; lane 2, mouse embryonic fibroblasts; lane 3, positive control (E14 es cells); lane 4, polymerase chain reaction control without the addition of template.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
In vitro differentiation of follicle-derived, homozygous embryonic stem (ES) cells (A) established by parthenogenetic activation and subsequent subculture of inner cell mass cell colonies in modified knockout Dulbecco's minimum essential medium supplemented with a 3:1 mixture of fetal bovine serum and knockout serum replacement. The colonies of follicle-derived ES cells were cultured in leukemia inhibitory factor-free medium for spontaneous differentiation into embryoid bodies. Immunocytochemistry of the embryoid bodies was conducted by using three germ layer-specific markers: neural cadherin adhesion molecule (for ectoderm; A), muscle actin (mesoderm; B), α-fetoprotein (endoderm; C), S-100 (ectoderm; D), desmin (mesoderm; E), and Troma-1 (endoderm; F). Cells consisting of embryoid bodies were positively stained with one of these markers. Bars = 50 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

6. Figure 5
Teratoma formation of follicle-derived, homozygous embryonic stem (ES) cells (A) established by parthenogenetic activation and subsequent subculture of inner cell mass cell colonies in modified knockout DMEM supplemented with a 3:1 mixture of fetal bovine serum and knockout serum replacement, 8 weeks after transplantation into NOD-SCID mice. The morphology of the teratoma was examined by hematoxylin and eosin staining of paraffin-embedded tissue. The teratoma contains a glandular stomachlike structure (A), exocrine pancreatic tissue (B), and respiratory epithelium with cilia (arrowhead, C), all derived from endodermal cells; stratified squamous epithelium with keratin (D), neuroepithelial rosette (E), pigmented retinal epithelium (F), and a sebaceous gland (G), from ectodermal cells; and adipocytes (arrowhead, H) and skeletal muscle bundles (arrow, H), from mesodermal cells. Bars = 200 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

7. Figure 6
Neuronal differentiation of preantral follicle-derived homozygous embryonic stem (ES) cells. (A, E) Phase-contrast images of differentiated follicle-derived, autologous ES cells in modified N2B27 medium. Tuj1-positive (B) and nestin-positive (C) neurons generated 7–10 days after replating on fibronectin. (D) Merged image of Tuj1-positive (B) and nestin-positive (C) neurons. Glial fibrillary acidic protein-positive astrocytes (F) and O4-positive oligodendrocytes (G) generated 11–14 days after replating on fibronectin, respectively. (H) Merged image of glial fibrillary acidic protein-positive astrocytes and O4-positive oligodendrocytes. Bars = 40 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions