Abstract
Human parthenogenetic stem cells are derived from the inner cell mass of blastocysts obtained from unfertilized oocytes that have been stimulated to develop without any participation of male gamete. As parthenogenesis does not involve the destruction of a viable human embryo, the derivation and use of human parthenogenetic stem cells does not raise the same ethical concerns as conventional embryonic stem cells. Human parthenogenetic stem cells are similar to embryonic stem cells in their proliferation and multilineage in vitro differentiation capacity. The aim of this study is to derive multipotent neural stem cells from human parthenogenetic stem cells that are stable to passaging and cryopreservation, and have the ability to further differentiate into functional neurons. Immunocytochemistry, quantitative real-time PCR, or FACS were used to confirm that the derived neural stem cells express neural markers such as NES, SOX2 and MS1. The derived neural stem cells keep uniform morphology for at least 30 passages and can be spontaneously differentiated into cells with neuron morphology that express TUBB3 and MAP2, and fire action potentials. These results suggest that parthenogenetic stem cells are a very promising and potentially unlimited source for the derivation of multipotent neural stem cells that can be used for therapeutic applications.
References
- 1 Revazova ES, Turovets NA, Kochetkova OD et al. HLA homozygous stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells10,11–24 (2008).
- 2 Revazova ES, Turovets NA, Kochetkova OD et al. Patient-specific stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells9,432–449 (2007).
- 3 Taylor CJ, Bolton EM, Pocock S et al. Banking on human embryonic stem cells: estimating the number of donor cell lines needed for HLA matching. Lancet366,2019–2025 (2005).
- 4 Reynolds BA, Weiss S. Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science225,1707–1710 (1992).
- 5 Okano H. Neural stem cells and strategies for the regeneration of the central nervous system. Proc. Jpn Acad. Ser. B Phys. Biol. Sci.86,438–450 (2010).
- 6 Eckardt S, Dinger TC, Kurosaka S et al.In vivo and in vitro differentiation of uniparental embryonic stem cells into hematopoietic and neural cell types. Organogenesis4,33–41 (2008).
- 7 Cibelli JB, Grant KA, Chapman KB et al. Parthenogenetic stem cells in nonhuman primates. Science295,819 (2002).
- 8 Harness JV, Turovets NA, Seiler MJ et al. Equivalence of conventionally-derived and parthenote-derived human embryonic stem cells. PLoS One6,e14499 (2011).
- 9 Nagy A, Sass M, Markkula M. Systematic non-uniform distribution of parthenogenetic cells in adult mouse chimaeras. Development106,321–324 (1989).
- 10 Sánchez-Pernaute R, Studer L, Ferrari D et al. Long-term survival of dopamine neurons derived from parthenogenetic primate embryonic stem cells (cyno-1) after transplantation. Stem Cells23(7),914–922 (2005).
- 11 Shin S, Mitalipova M, Noggle S et al. Long-term proliferation of human embryonic stem cell-derived neuroepithelial cells using defined adherent culture conditions. Stem Cells24,125–138 (2006).
- 12 Dhara SK, Stice SL. Neural differentiation of human embryonic stem cells. J. Cell. Biochem.105,633–640 (2008).
- 13 Archer TC, Jin J, Casey ES. Interaction of Sox1, Sox2, Sox3 and Oct4 during primary neurogenesis. Dev. Biol.350,429–440 (2011).
- 14 Colleoni S, Galli C, Giannelli SG et al. Long-term culture and differentiation of CNS precursors derived from anterior human neural rosettes following exposure to ventralizing factors. Exp. Cell. Res.316,1148–1158 (2010).
- 15 Graham V, Khudyakov J, Ellis P, Pevny L. SOX2 functions to maintain neural progenitor identity. Neuron39,749–765 (2003).
