Establishment of a Human Embryonic Germ Cell Line and Comparison with Mouse and Human Embryonic Stem Cells

다국어 초록 (Multilingual Abstract)

Human embryonic stem (ES) cells and embryonic germ (EG) cells are pluripotent and are invaluable material for in vitro studies of human embryogenesis and cell therapy. So far, only two groups have reported the establishment of human EG cell lines, whereas at least five human ES cell lines have been established. To see if human EG cell lines can be reproducibly estab-lished, we isolated primordial germ cells (PGCs) from gonadal ridges and mesenteries (9 weeks post-fertilization) and cultured them on mouse STO cells. As with mouse ES colonies, the PGC-derived cells have given rise to multilayered colonies without any differ-entiation over a year of continuous culture. They are karyotypically normal and express high levels of alka-line phosphatase, Oct-4, and several cell-surface mark-ers. Histological and immunocytochemical analysis of embryoid bodies (EBs) formed from floating cultures of the PGC-derived cell colonies revealed ectodermal, endodermal, and mesodermal tissues. When the EBs were cultured in the presence of insulin, transferrin, sodium selenite, and fibronectin for 1 week, markers of primitive neuroectoderm were expressed in cells within the EBs as well as in cells growing out from the EBs. These observations indicate that our PGC-derived cells satisfy the criteria for pluripotent stem cells and hence may be EG cells.

참고문헌 (Reference)

1 Billinghurst, "evidence from the dysmyelinated shiverer mouse brain" 96 : 70297034-, 1999

2 McDonald, "differentiate and promote recovery in injured rat spinal cord" 5 : 1410-1412, 1999

3 A, "and migration of the primordial germ cells in the mouse embryo" 1954

4 Wechsler, "Transplantation of cultured human neuronal cells for patients with stroke" 55 : 565569-, 2000

5 Katz, "The in vitro development of blastocyst-derived embryonic stem cell lines formation of visceral yolk sac" 1985

6 Stevens, "Studies on transplantable testicular teratomas of strain 129 mice" 1958

7 Thomson, "Pluripotent cell lines derived from common marmoset" 55 : 254259. -, 1996

8 Snyder, "Neural progenitor cell engraftment corrects lysosomal storage throughout the MPS VII mouse brain." 374 : 367370.-, 1995

9 Williams, "Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells" -336, 1988

10 Resnick, "Long-term proliferation of mouse primordial germ cells in culture" -359, 1992

11 Pain, "Long-term in vitro culture and characterization of avian embryonic stem cells with multiple morphogenetic potentialities" 122 : 23392348-, 1996

12 Kim, "Knock-out effect of transcription factor GATA-1 on early erythropoiesis" 6 : 176182-, 1996

13 Shim, "Isolation of pluripotent stem cells from cultured porcine primordial germ cells" 57 : 10891095-, 1997

14 Thomson, "Isolation of a primate embryonic stem cell line" 92 : 78447848-, 1995

15 Martin, "Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells" -78, 1981

16 Liu, "Intraspinal delivery of neurotrophin-3 using neural stem cells genetically modified by recombinant retrovirus" 158 : 926-, 1999

17 Zhang, "In vitro differentiation of transplantable neural precursors from human embryonic stem cells" 19 : 1129-1133, 2001

18 Zhang, "In vitro differentiation of transplantable neural precursors from human embryonic stem cells" 19 : 1129-1133, 2001

19 I, "Immunohistochemical localization of murine stage-specific embryonic antigens in human testicular germ cell tumors" damja (damja): 1982

20 Calhoun, "Human embryonic stem cell lines derived from discarded embryos" 21 : 521526-, 2003

21 Kerr, "Human embryonic germ cell derivatives facilitate motor recovery of rats with diffuse motor neuron injury" 23 : 51315140-, 2003

22 Andrews, "Human embryonal carcinoma cells and their differentiation in culture" 1987

23 Cheng, "Human adult marrow cells support prolonged expansion of human embryonic stem cells in culture" 21 : 131142-, 2003

24 Benedetti, "Gene therapy of experimental brain tumors using neural progenitor cells" 6 : 447450.-, 2000

25 Lacorazza, H. D.,, "Expression of human beta-hexosaminidase alpha-subunit gene the gene defect of Tay-Sachs disease in mouse brains upon engraftment of transduced progenitor cells" 2 : 424429-, 1996

26 OEiges, R.,, "Establishment of human embryonic stem cell-transfected clones carrying a marker for undifferentiated cells" 11 : 514518.-, 2001

27 Williams, "Establishment of hamster blastocyst-derived embryonic stem" 1988

28 Evans, "Establishment in culture of pluripotential cells from mouse embryos and DAX1 expression patterns during human sex determination and gonadal development" -292, 19812000

29 Park, "Establishment and maintenance of human embryonic stem cells on STO [Epub ahead of print]" 113 (113): 510-, 20032000

30 A, "Embryonic stem cells derived from morulae and blastocysts of mink comparisons of their pluripotencies" seme (seme): 1993

31 Pera, "Embryonic stem cell lines from human blastocysts somatic differentiation in vitro" 18 (18): 399404.-, 2000

32 Thomson, "Embryonic stem cell lines derived from human blastocysts" 282 : 11451147-, 1998

33 Sung, "Differentiation and apoptosis of the mammalian embryo and embryonic stem cells Establishment of mouse ESC and induction of differentiation by reproductive hormones" 6 : 5566.-, 2002

34 Wang, "Derivation of pluripotent stem cells from cultured human primordial germ cells" 95 : 1372613731-, 1998

35 Turnpenny, L, "Derivation of human embryonic germ cells an alternative source of pluripotent stem cells" 21 : 598609-, 2003

36 A, "Analysis of lymphokine mRNA expression and relevance to cancer immunotherapy" immu (immu): 1989

37 Cannon, "Altered differentiation of CNS neural progenitor cells after transplantation into the injured adult rat spinal cord" 6 : 327338-, 1997