Skip to main content
SpringerLink
Log in

Derivation of Human Embryonic Stem Cells (hESC)

  • Protocol
  • First Online:
Human Fertility

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1154))

  • 4026 Accesses

Abstract

Stem cells are characterized by their absolute or relative lack of specialization their ability for self-renewal, as well as their ability to generate differentiated progeny through cellular lineages with one or more branches. The increased availability of embryonic tissue and greatly improved derivation methods have led to a large increase in the number of hESC lines.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ramalho-Santos M, Willenbring H (2007) On the origin of the term “stem cell”. Cell Stem Cell 1:35–38

    PubMed  CAS  Google Scholar 

  2. Martin GR, Evans MJ (1974) The morphology and growth of a pluripotent teratocarcinoma cell line and its derivatives in tissue culture. Cell 2:163–172

    PubMed  CAS  Google Scholar 

  3. Stevens LC (1970) The development of transplantable teratocarcinomas from intratesticular grafts of pre- and postimplantation mouse embryos. Dev Biol 21:364–382

    PubMed  CAS  Google Scholar 

  4. Martin GR, Evans MJ (1975) Differentiation of clonal lines of teratocarcinoma cells: formation of embryoid bodies in vitro. Proc Natl Acad Sci U S A 72:1441–1445

    PubMed Central  PubMed  CAS  Google Scholar 

  5. Andrews PW (2002) From teratocarcinomas to embryonic stem cells. Philos Trans R Soc Lond B Biol Sci 357:405–417

    PubMed Central  PubMed  Google Scholar 

  6. Andrews PW (1984) Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol 103:285–293

    PubMed  CAS  Google Scholar 

  7. Solter D (2006) From teratocarcinomas to embryonic stem cells and beyond: a history of embryonic stem cell research. Nat Rev Genet 7:319–327

    PubMed  CAS  Google Scholar 

  8. Solter D (2005) What is a stem cell? Novartis Found Symp 265(3–12):92–97

    Google Scholar 

  9. Edwards RG (2004) In: Lanza R et al (eds) Handbook of stem cells. Elsevier Academic Press, Burlington, MA, pp 1–14

    Google Scholar 

  10. Evans M, Kaufman M (1981) Establishment in culture of pluripotent cells from mouse embryos. Nature 292:154–156

    PubMed  CAS  Google Scholar 

  11. Martin G (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci U S A 78:7635

    Google Scholar 

  12. Brons IG et al (2007) Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature. doi:10.1038/05950

    PubMed  Google Scholar 

  13. Tesar PJ et al (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature. doi:10.038/05972

    PubMed  Google Scholar 

  14. Nichols J, Smith A (2011) The origin and identity of embryonic stem cells. Development 138:3–8

    PubMed  CAS  Google Scholar 

  15. Nicholds J, Smith A (2009) Naïve and primed pluripotent states. Cell Stem Cell 4:487–492

    Google Scholar 

  16. Resnick JL et al (1992) Long-term proliferation of mouse primordial germ cells in culture. Nature 359:550–551

    PubMed  CAS  Google Scholar 

  17. Thomson JA et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147

    PubMed  CAS  Google Scholar 

  18. Shamblott MJ et al (1998) Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc Natl Acad Sci U S A 95:13726–13731

    PubMed Central  PubMed  CAS  Google Scholar 

  19. Gerecht-Nir S, Itskovitz-Eldor J (2004) The promise of human embryonic stem cells. Best Pract Res Clin Obstet Gynaecol 18:843–852

    PubMed  Google Scholar 

  20. Gerecht-Nir S, Itskovitz-Eldor J (2004) Cell therapy using human embryonic stem cells. Transpl Immunol 12:203–209

    PubMed  CAS  Google Scholar 

  21. Golan-Mashiach M et al (2005) Design principle of gene expression used by human stem cells: implication for pluripotency. FASEB J 19:147–149

    PubMed  CAS  Google Scholar 

  22. Veeck L, Zaninovic N (eds) (2003) An atlas of human blastocyst. Parthenon, New York

    Google Scholar 

  23. Edwards RG (2005) Changing genetic world of IVF, stem cells and PGD. B. Polarities and gene expression in differentiating embryo cells and stem cells. Reprod Biomed Online 1:761–776

    Google Scholar 

  24. Niwa H et al (2005) Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell 123:917–929

    PubMed  CAS  Google Scholar 

  25. Heins N et al (2006) Clonal derivation and characterization of human embryonic stem cell lines. J Biotechnol 122:511–520

    PubMed  CAS  Google Scholar 

  26. Palmieri SL et al (1994) Oct-4 transcription factor is differentially expressed in the mouse embryo during establishment of the first two extraembryonic cell lineages involved in implantation. Dev Biol 166:259–267

    PubMed  CAS  Google Scholar 

  27. Huntriss J et al (2004) Expression of mRNAs for DNA methyltransferases and methyl-CpG-binding proteins in the human female germ line, preimplantation embryos, and embryonic stem cells. Mol Reprod Dev 67:323–336

    PubMed  CAS  Google Scholar 

  28. Cauffman G et al (2005) Oct-4 mRNA and protein expression during human preimplantation development. Mol Hum Reprod 11:173–181

    PubMed  CAS  Google Scholar 

  29. Hyslop L et al (2005) Downregulation of NANOG induces differentiation of human embryonic stem cells to extraembryonic lineages. Stem Cells 23:1035–1043

    PubMed  CAS  Google Scholar 

  30. Zhang X et al (2006) Derivation of human embryonic stem cells from developing and arrested embryos. Stem Cells 24:2669–2676

    PubMed  CAS  Google Scholar 

  31. Adjaye J et al (2005) Primary differentiation in the human blastocyst: comparative molecular portraits of inner cell mass and trophectoderm cells. Stem Cells 23:1514–1525

    PubMed  CAS  Google Scholar 

  32. Stephenson EL et al (2006) Proposal for a universal minimum information convention for the reporting on the derivation of human embryonic stem cell lines. Regen Med 1:739–750

    PubMed  Google Scholar 

  33. Amit M, Itskovitz-Eldor J (2006) Sources, derivation, and culture of human embryonic stem cells. Semin Reprod Med 24:298–303

    PubMed  CAS  Google Scholar 

  34. Solter D, Knowles BB (1975) Immunosurgery of mouse blastocyst. Proc Natl Acad Sci U S A 72:5099–5102

    PubMed Central  PubMed  CAS  Google Scholar 

  35. Moon S et al (2006) Generation, culture, and differentiation of human embryonic stem cells for therapeutic applications. Mol Ther 13:5–14

    PubMed  CAS  Google Scholar 

  36. Kim HS et al (2005) Methods for derivation of human embryonic stem cells. Stem Cells 23:1228–1233

    PubMed  Google Scholar 

  37. Amit M, Itskovitz-Eldor J (2002) Derivation and spontaneous differentiation of human embryonic stem cells. J Anat 200:225–232

    PubMed Central  PubMed  Google Scholar 

  38. Turetsky T et al (2008) Laser-assisted derivation of human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis. Hum Reprod 23:46–53

    PubMed  CAS  Google Scholar 

  39. Chen AE et al (2009) Optimal timing of inner cell mass isolation increases the efficiency of human embryonic stem cell derivation and allows generation of sibling cell lines. Cell Stem Cell 4:103–106

    PubMed Central  PubMed  CAS  Google Scholar 

  40. Zaninovic N et al (2010) Efficiency of hESC derivation: optimization of the ICM isolation and culture conditions. ASRM, Denver, Colorado

    Google Scholar 

  41. Heins N et al (2004) Derivation, characterization, and differentiation of human embryonic stem cells. Stem Cells 22:367–376

    PubMed  Google Scholar 

  42. Reubinoff BE et al (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18:399–404

    PubMed  CAS  Google Scholar 

  43. Mitalipova M et al (2003) Human embryonic stem cell lines derived from discarded embryos. Stem Cells 21:521–526

    PubMed  CAS  Google Scholar 

  44. Chen H et al (2005) The derivation of two additional human embryonic stem cell lines from day 3 embryos with low morphological scores. Hum Reprod 20:2201–2206

    PubMed  Google Scholar 

  45. Levron J et al (1995) Male and female genomes associated in a single pronucleus in human zygotes. Biol Reprod 52:653–657

    PubMed  CAS  Google Scholar 

  46. Suss-Toby E et al (2004) Derivation of a diploid human embryonic stem cell line from a mononuclear zygote. Hum Reprod 19:670–675

    PubMed  Google Scholar 

  47. Cibelli J et al (2006) Embryonic stem cells from parthenotes. Methods Enzymol 418:117–135

    PubMed  CAS  Google Scholar 

  48. Revazova ES et al (2007) Patient-specific stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells 9:432–449

    PubMed  CAS  Google Scholar 

  49. Plachot M (1992) Cytogenetic analysis of oocytes and embryos. Ann Acad Med Singapore 21:538–544

    PubMed  CAS  Google Scholar 

  50. Sandalinas M et al (2001) Developmental ability of chromosomally abnormal human embryos to develop to the blastocyst stage. Hum Reprod 16:1954–1958

    PubMed  CAS  Google Scholar 

  51. Baharvand H et al (2006) Generation of new human embryonic stem cell lines with diploid and triploid karyotypes. Dev Growth Differ 48:117–128

    PubMed  Google Scholar 

  52. Veeck LL et al (2004) High pregnancy rates can be achieved after freezing and thawing human blastocysts. Fertil Steril 82:1418–1427

    PubMed  Google Scholar 

  53. Sjogren A et al (2004) Human blastocysts for the development of embryonic stem cells. Reprod Biomed Online 9:326–329

    PubMed  Google Scholar 

  54. Tesar PJ (2005) Derivation of germ-line-competent embryonic stem cell lines from preblastocyst mouse embryos. Proc Natl Acad Sci U S A 102:8239–8244

    PubMed Central  PubMed  CAS  Google Scholar 

  55. Chung Y et al (2006) Embryonic and extraembryonic stem cell lines derived from single mouse blastomeres. Nature 439:216–219

    PubMed  CAS  Google Scholar 

  56. Fong CY et al (2006) Unsuccessful derivation of human embryonic stem cell lines from pairs of human blastomeres. Reprod Biomed Online 13:295–300

    PubMed  Google Scholar 

  57. Lerou PH et al (2008) Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryo. Nat Protoc 3:923–933

    PubMed  CAS  Google Scholar 

  58. Stojkovic M et al (2004) Derivation of human embryonic stem cells from day-8 blastocysts recovered after three-step in vitro culture. Stem Cells 22:790–797

    PubMed  Google Scholar 

  59. Markert CL, Petters RM (1978) Manufactured hexaparental mice show that adults are derived from three embyronic cells. Science 202:56–58

    PubMed  CAS  Google Scholar 

  60. Strelchenko N et al (2004) Morula-derived human embryonic stem cells. Reprod Biomed Online 9:623–629

    PubMed  Google Scholar 

  61. Solter D (2005) Politically correct human embryonic stem cells? N Engl J Med 353:2321–2323

    PubMed  CAS  Google Scholar 

  62. Wakayama S et al (2007) Efficient establishment of mouse embryonic stem cell lines from single blastomeres and polar bodies. Stem Cells 25:986–993

    PubMed  CAS  Google Scholar 

  63. Klimanskaya I et al (2006) Human embryonic stem cell lines derived from single blastomeres. Nature 444(7118):481–485

    PubMed  CAS  Google Scholar 

  64. Chung Y et al (2008) Human embryonic stem cell lines generated without embryo destruction. Cell Stem Cell. doi:10.1016/j.stem.2007.12.013

    PubMed Central  Google Scholar 

  65. Ilic D et al (2009) Derivation of human embryonic stem cell lines from biopsied blastomeres on human feeders with minimal exposure to xenomaterials. Stem Cells Dev 18:1343–1350

    PubMed  CAS  Google Scholar 

  66. Geens M et al (2009) Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos. Hum Reprod 24:2709–2717

    PubMed Central  PubMed  CAS  Google Scholar 

  67. Hao J et al (2007) Derivation of mouse embryonic stem cells (mESC) from individual and aggregated mouse blastomeres. ASRM, Washington DC

    Google Scholar 

  68. Munné S et al (2005) Self-correction of chromosomally abnormal embryos in culture and implications for stem cell production. Fertil Steril 84:1328–1334

    PubMed  Google Scholar 

  69. Los F et al (1998) Uniparental disomy with and without confined placental mosaicism: a model for trisomic zygote rescue. Prenat Diagn 18:659–668

    PubMed  CAS  Google Scholar 

  70. Shaffer LG et al (2001) American College of Medical Genetics statement of diagnostic testing for uniparental disomy. Genet Med 3:206–211

    PubMed Central  PubMed  CAS  Google Scholar 

  71. Verlinsky Y et al (2006) Repository of human embryonic stem cell lines and development of individual specific lines using stembrid technology. Reprod Biomed Online 13:547–550

    PubMed  CAS  Google Scholar 

  72. Verlinsky Y et al (2005) Human embryonic stem cell lines with genetic disorders. Reprod Biomed Online 10:105–110

    PubMed  CAS  Google Scholar 

  73. Mateizel I et al (2006) Derivation of human embryonic stem cell lines from embryos obtained after IVF and after PGD for monogenic disorders. Hum Reprod 21:503–511

    PubMed  CAS  Google Scholar 

  74. Löser P et al (2010) Human embryonic stem cell lines and their use in international research. Stem Cells 28:240–246

    PubMed Central  PubMed  Google Scholar 

  75. Ährlund-Richter L et al (2009) Isolation and production of cells suitable for human therapy: challenges ahead. Cell Stem Cell 4:20–26

    PubMed  Google Scholar 

  76. Hasegawa K et al (2010) Current technology for the derivation of pluripotent stem cell lines from human embryos. Cell Stem Cell 6:521–531

    PubMed  CAS  Google Scholar 

  77. Hoffman LM, Carpenter MK (2005) Characterization and culture of human embryonic stem cells. Nat Biotechnol 23:699–708

    PubMed  CAS  Google Scholar 

  78. Crook JM et al (2007) The generation of six clinical-grade human embryonic stem cell lines. Cell Stem Cell 1:490–494

    CAS  Google Scholar 

  79. Ellerström C et al (2006) Derivation of a xeno-free human embryonic stem cell line. Stem Cells 24:2170–2176

    PubMed  Google Scholar 

  80. Rajala K et al (2007) Testing of nine different xeno-free culture media for human embryonic stem cell cultures. Hum Reprod 22:1231–1238

    PubMed  CAS  Google Scholar 

  81. Williams RL et al (1988) Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336:684–687

    PubMed  CAS  Google Scholar 

  82. Smith AG et al (1988) Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336:688–690

    PubMed  CAS  Google Scholar 

  83. Niwa H et al (1998) Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev 12:2048–2060

    PubMed Central  PubMed  CAS  Google Scholar 

  84. Humphrey RK et al (2004) Maintenance of pluripotency in human embryonic stem cells is STAT3 independent. Stem Cells 22:522–530

    PubMed  CAS  Google Scholar 

  85. Sato N et al (2003) Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat Med 10(1):55–63

    PubMed  Google Scholar 

  86. Levenstein ME et al (2006) Basic fibroblast growth factor support of human embryonic stem cell self-renewal. Stem Cells 24:568–574

    PubMed  CAS  Google Scholar 

  87. James D et al (2005) TGFbeta/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells. Development 132:1273–1282

    PubMed  CAS  Google Scholar 

  88. Amit M et al (2000) Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev Biol 2000(227):271–278

    Google Scholar 

  89. Park SP et al (2004) Establishment of human embryonic stem cell lines from frozen-thawed blastocysts using STO cell feeder layers. Hum Reprod 19:676–684

    PubMed  Google Scholar 

  90. Choo A et al (2006) Immortalized feeders for the scale-up of human embryonic stem cells in feeder and feeder-free conditions. J Biotechnol 122:130–141

    PubMed  CAS  Google Scholar 

  91. Martin MJ et al (2005) Human embryonic stem cells express an immunogenic nonhuman sialic acid. Nat Med 11:228–232

    PubMed  CAS  Google Scholar 

  92. Amit M et al (2005) No evidence for infection of human embryonic stem cells by feeder cell-derived murine leukemia viruses. Stem Cells 23:761–771

    PubMed  CAS  Google Scholar 

  93. Skottman H, Hovatta O (2006) Culture conditions for human embryonic stem cells. Reproduction 132:691–698

    PubMed  CAS  Google Scholar 

  94. Mallon BS et al (2006) Toward xeno-free culture of human embryonic stem cells. Int J Biochem Cell Biol 38:1063–1075

    PubMed Central  PubMed  CAS  Google Scholar 

  95. Amit M et al (2003) Human feeder layers for human embryonic stem cells. Biol Reprod 68:2150–2156

    PubMed  CAS  Google Scholar 

  96. Richards M et al (2003) Comparative evaluation of various human feeders for prolonged undifferentiated growth of human embryonic stem cells. Stem Cells 21:546–556

    PubMed  CAS  Google Scholar 

  97. Cheng L et al (2003) Human adult marrow cells support prolonged expansion of human embryonic stem cells in culture. Stem Cells 21:131–142

    PubMed  CAS  Google Scholar 

  98. Rosler ES et al (2004) Long-term culture of human embryonic stem cells in feeder-free conditions. Dev Dyn 229:259–274

    PubMed  CAS  Google Scholar 

  99. Stojkovic P et al (2005) An autogeneic feeder cell system that efficiently supports growth of undifferentiated human embryonic stem cells. Stem Cells 23:306–314

    PubMed  CAS  Google Scholar 

  100. Hovatta O et al (2003) A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells. Hum Reprod 18:1404–1409

    PubMed  Google Scholar 

  101. Genbacev O et al (2005) Serum-free derivation of human embryonic stem cell lines on human placental fibroblast feeders. Fertil Steril 83:1517–1529

    PubMed  Google Scholar 

  102. Lee JB et al (2005) Establishment and maintenance of human embryonic stem cell lines on human feeder cells derived from uterine endometrium under serum-free condition. Biol Reprod 72:42–49

    PubMed  CAS  Google Scholar 

  103. Xu C et al (2004) Immortalized fibroblast-like cells derived from human embryonic stem cells support undifferentiated cell growth. Stem Cells 22:972–980

    PubMed  CAS  Google Scholar 

  104. Richards M et al (2002) Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells. Nat Biotechnol 20:933–936

    PubMed  CAS  Google Scholar 

  105. Wang Q et al (2005) Derivation and growing human embryonic stem cells on feeders derived from themselves. Stem Cells 23:1221–1227

    PubMed  Google Scholar 

  106. Amit M et al (2004) Feeder layer- and serum-free culture of human embryonic stem cells. Biol Reprod 70:837–845

    PubMed  CAS  Google Scholar 

  107. Koivisto H et al (2004) Cultures of human embryonic stem cells: serum replacement medium or serum-containing media and the effect of basic fibroblast growth factor. Reprod Biomed Online 9:330–337

    PubMed  CAS  Google Scholar 

  108. Carpenter MK et al (2004) Properties of four human embryonic stem cell lines maintained in a feeder-free culture system. Dev Dyn 229:243–258

    PubMed  CAS  Google Scholar 

  109. Xu C et al (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat Biotechnol 19:971–974

    PubMed  CAS  Google Scholar 

  110. Carpenter M et al (2003) Characterization and differentiation of human embryonic stem cells. Cloning Stem Cells 5:79–88

    PubMed  CAS  Google Scholar 

  111. Kleinman HK et al (1982) Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21:6188–6193

    PubMed  CAS  Google Scholar 

  112. Xu RH et al (2005) Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. Nat Methods 2:185–190

    PubMed  CAS  Google Scholar 

  113. Xu C et al (2005) Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. Stem Cells 23:315–323

    PubMed  CAS  Google Scholar 

  114. Wang G et al (2005) Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers. Biochem Biophys Res Commun 330:934–942

    PubMed  CAS  Google Scholar 

  115. Li Y et al (2005) Expansion of human embryonic stem cells in defined serum-free medium devoid of animal-derived products. Biotechnol Bioeng 91:688–698

    PubMed  CAS  Google Scholar 

  116. Ludwig TE et al (2006) Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24:185–187

    PubMed  CAS  Google Scholar 

  117. Stojkovic P et al (2005) Human-serum matrix supports undifferentiated growth of human embryonic stem cells. Stem Cells 23:895–902

    PubMed  CAS  Google Scholar 

  118. Peiffer I et al (2010) Optimization of physiological xenofree molecularly defined media and matrices to maintain human embryonic stem cell pluripotency. Methods Mol Biol 584:97–108

    PubMed  CAS  Google Scholar 

  119. Wagner KE, Vemuri MC (2010) Serum-free and feeder-free culture expansion of human embryonic stem cells. Methods Mol Biol 584:109–119

    PubMed  Google Scholar 

  120. Chen G et al (2011) Chemically defined conditions for human iPSC derivation and culture. Nat Protoc 8:424–429

    CAS  Google Scholar 

  121. Yim EK, Leong KW (2005) Proliferation and differentiation of human embryonic germ cell derivatives in bioactive polymeric fibrous scaffold. J Biomater Sci Polym Ed 16:1193–1217

    PubMed  CAS  Google Scholar 

  122. Kroupova J et al (2006) Functional polymer hydrogels for embryonic stem cell support. J Biomed Mater Res B Appl Biomater 76:315–325

    PubMed  Google Scholar 

  123. Elisseeff J et al (2005) Advances in skeletal tissue engineering with hydrogels. Orthod Craniofac Res 8:150–161

    PubMed  CAS  Google Scholar 

  124. Inzunza J et al (2005) Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells. Stem Cells 23:544–549

    PubMed  CAS  Google Scholar 

  125. Hong-mei P, Gui-an C (2006) Serum-free medium cultivation to improve efficacy in establishment of human embryonic stem cell lines. Hum Reprod 21:217–222

    PubMed  Google Scholar 

  126. Chen HF et al (2007) Derivation, characterization and differentiation of human embryonic stem cells: comparing serum-containing versus serum-free media and evidence of germ cell differentiation. Hum Reprod 22:567–577

    PubMed  Google Scholar 

  127. Klimanskaya I et al (2005) Human embryonic stem cells derived without feeder cells. Lancet 365:1636–1641

    PubMed  CAS  Google Scholar 

  128. Draper JS et al (2004) Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol 22:53–54

    PubMed  CAS  Google Scholar 

  129. Fletcher JM et al (2006) Variations in humanized and defined culture conditions supporting derivation of new human embryonic stem cell lines. Cloning Stem Cells 8:319–334

    PubMed  CAS  Google Scholar 

  130. Prowse AB et al (2005) A proteome analysis of conditioned media from human neonatal fibroblasts used in the maintenance of human embryonic stem cells. Proteomics 5:978–989

    PubMed  CAS  Google Scholar 

  131. Chin AC et al (2007) Identification of proteins from feeder conditioned medium that support human embryonic stem cells. J Biotechnol 130:320–328

    PubMed  CAS  Google Scholar 

  132. Lim JW, Bodnar A (2002) Proteome analysis of conditioned medium from mouse embryonic fibroblast feeder layers which support the growth of human embryonic stem cells. Proteomics 2:1187–1203

    PubMed  CAS  Google Scholar 

  133. Stewart MH et al (2006) Clonal isolation of hESCs reveals heterogeneity within the pluripotent stem cell compartment. Nat Methods 3:807–815

    PubMed  CAS  Google Scholar 

  134. Lauss M et al (2005) Single inner cell masses yield embryonic stem cell lines differing in lifr expression and their developmental potential. Biochem Biophys Res Commun 331:1577–1586

    PubMed  CAS  Google Scholar 

  135. Hewitt Z et al (2006) Fluorescence-activated single cell sorting of human embryonic stem cells. Cloning Stem Cells 8:225–234

    PubMed  CAS  Google Scholar 

  136. Sidhu KS, Tuch BE (2006) Derivation of three clones from human embryonic stem cell lines by FACS sorting and their characterization. Stem Cells Dev 15:61–69

    PubMed  CAS  Google Scholar 

  137. Wong RC et al (2006) Gap junctions modulate apoptosis and colony growth of human embryonic stem cells maintained in a serum-free system. Biochem Biophys Res Commun 344:181–188

    PubMed  CAS  Google Scholar 

  138. Forsyth NR et al (2006) Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities. Cloning Stem Cells 8:16–23

    PubMed  CAS  Google Scholar 

  139. Damoiseaux R et al (2009) Integrated chemical genomics reveals modifiers of survival in human embryonic stem cells. Stem Cells 27:533–542

    PubMed Central  PubMed  CAS  Google Scholar 

  140. Watanabe K et al (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25:681–686

    PubMed  CAS  Google Scholar 

  141. Zweigerdt R et al (2011) Scalable expansion of human pluripotent stem cells in suspension culture. Nat Protoc 6:689–700

    PubMed  CAS  Google Scholar 

  142. Amit M et al (2010) Suspension culture of undifferentiated human embryonic and induced pluripotent stem cells. Stem Cell Rev Rep 6:248–259

    Google Scholar 

  143. Steiner D et al (2010) Derivation, propagation and controlled differentiation of human embryonic stem cells in suspension. Nat Biotechnol 28:361–364

    PubMed  CAS  Google Scholar 

  144. Krawetz R et al (2010) Large-scale expansion of pluripotent human embryonic stem cells in stirred-suspension bioreactors. Tissue Eng Part C Methods 16:573–582

    PubMed  CAS  Google Scholar 

  145. Singh H et al (2010) Up-scaling single cell-inoculated suspension culture of human embryonic stem cells. Stem Cell Res 4:165–179

    PubMed  CAS  Google Scholar 

  146. Oh SKW et al (2009) Long-term microcarrier suspension cultures of human embryonic stem cells. Stem Cell Res 2:219–230

    PubMed  CAS  Google Scholar 

  147. Chen AKL et al (2011) Critical microcarrier properties affecting the expansion of undifferentiated human embryonic stem cells. Stem Cell Res 7:97–111

    PubMed  CAS  Google Scholar 

  148. Thomas RJ et al (2009) Automated, scalable culture of human embryonic stem cells in feeder-free conditions. Biotechnol Bioeng 102:1636–1644

    PubMed  CAS  Google Scholar 

  149. Baker DE et al (2007) Adaptation to culture of human embryonic stem cells and oncogenesis in vivo. Nat Biotechnol 25:207–215

    PubMed  CAS  Google Scholar 

  150. Longo L et al (1997) The chromosome make-up of mouse embryonic stem cells is predictive of somatic and germ cell chimaerism. Transgenic Res 6:321–328

    PubMed  CAS  Google Scholar 

  151. Liu X et al (1997) Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev Dyn 209:85–91

    PubMed  CAS  Google Scholar 

  152. Inzunza J et al (2004) Comparative genomic hybridization and karyotyping of human embryonic stem cells reveals the occurrence of an isodicentric X chromosome after long-term cultivation. Mol Hum Reprod 10:461–466

    PubMed  CAS  Google Scholar 

  153. Wilton L (2002) Preimplantation genetic diagnosis for aneuploidy screening in early human embryos: a review. Prenat Diagn 22:512–518

    PubMed  Google Scholar 

  154. Atkin NB, Baker MC (1982) Specific chromosome change, i(12p), in testicular tumours? Lancet 2:1349

    PubMed  CAS  Google Scholar 

  155. Skotheim RI et al (2002) New insights into testicular germ cell tumorigenesis from gene expression profiling. Cancer Res 62:2359–2364

    PubMed  CAS  Google Scholar 

  156. Korkola JE et al (2006) Down-regulation of stem cell genes, including those in a 200-kb gene cluster at 12p13.31, is associated with in vivo differentiation of human male germ cell tumors. Cancer Res 66:820–827

    PubMed  CAS  Google Scholar 

  157. Mitalipova MM et al (2005) Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol 23:19–20

    PubMed  CAS  Google Scholar 

  158. Catalina P et al (2008) Human ESCs predisposition to karyotypic instability: is a matter of culture adaptation or differential vulnerability among hESC lines due to inherent properties? Mol Cancer 7:76

    PubMed Central  PubMed  Google Scholar 

  159. Ezashi T et al (2005) Low O2 tensions and the prevention of differentiation of hES cells. Proc Natl Acad Sci U S A 102:4783–4788

    PubMed Central  PubMed  CAS  Google Scholar 

  160. Brivanlou AH et al (2003) Stem cells. Setting standards for human embryonic stem cells. Science 300:913–916

    PubMed  CAS  Google Scholar 

  161. Adewumi O et al (2007) Characterization of human embryonic stem cell lines by the International Stem Cell Initiative. Nat Biotechnol 25:803–816

    PubMed  CAS  Google Scholar 

  162. Spagnoli FM, Hemmati-Brivanlou A (2006) Guiding embryonic stem cells towards differentiation: lessons from molecular embryology. Curr Opin Genet Dev 16(5):469–475

    PubMed  CAS  Google Scholar 

  163. Lawrenz B et al (2004) Highly sensitive biosafety model for stem-cell-derived grafts. Cytotherapy 6:212–222

    PubMed  CAS  Google Scholar 

  164. Niwa HJ et al (2000) Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet 24:372–376

    PubMed  CAS  Google Scholar 

  165. Chambers I et al (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113:643–655

    PubMed  CAS  Google Scholar 

  166. Boyer LA et al (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122:947–956

    PubMed Central  PubMed  CAS  Google Scholar 

  167. Babaie Y et al (2007) Analysis of Oct-4-dependent transcriptional networks regulating self-renewal and pluripotency in human embryonic stem cells. Stem Cells 25:500–510

    PubMed  CAS  Google Scholar 

  168. Boyer LA et al (2006) Molecular control of pluripotency. Curr Opin Genet Dev 16:455–462

    PubMed  CAS  Google Scholar 

  169. Tang F et al (2010) Tracing the derivation of embryonic stem cells from the inner cell mass by single-cell RNA-Seq analysis. Cell Stem Cell 6:468–478

    PubMed Central  PubMed  CAS  Google Scholar 

  170. Lavon N et al (2008) Derivation of euploid human embryonic stem cells from aneuploid embryos. Stem Cells 26:1874–1882

    PubMed  CAS  Google Scholar 

  171. Biancotti JC et al (2010) Human embryonic stem cells as models for aneuploid chromosomal syndromes. Stem Cells 28:1530–1540

    PubMed  CAS  Google Scholar 

  172. Zaninovic N et al (2008) “Self-correction” by uniparental disomy (UPD) of chromosomally abnormal embryos does not occur in human embryonic stem cell (hESC) cultures. ESHRE 2008, Barcelona, Spain

    Google Scholar 

  173. James D et al (2010) Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFbeta inhibition is Id1 dependent. Nat Biotechnol 28:161–166

    PubMed Central  PubMed  CAS  Google Scholar 

  174. James D et al (2011) Lentiviral transduction and clonal selection of hESCs with endothelial-specific transgenic reporters. Curr Protoc Stem Cell Biol Chapter 1:Unit1F.12

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Center for Reproductive Medicine, Weill Cornell Medical College, 1305 York Avenue, New York, NY, 10021, USA

    Nikica Zaninovic M.D., Qiansheng Zhan Ph.D. & Zev Rosenwaks M.D., Ph.D.

Authors
  1. Nikica Zaninovic M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qiansheng Zhan Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zev Rosenwaks M.D., Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. The Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical College, New York, New York, USA

    Zev Rosenwaks

  2. Dept. Developmental & Regenerative Biology, Ichan School of Medicine at Mount Sinai, New York, New York, USA

    Paul M. Wassarman

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Zaninovic, N., Zhan, Q., Rosenwaks, Z. (2014). Derivation of Human Embryonic Stem Cells (hESC). In: Rosenwaks, Z., Wassarman, P. (eds) Human Fertility. Methods in Molecular Biology, vol 1154. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0659-8_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0659-8_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0658-1

  • Online ISBN: 978-1-4939-0659-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation