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Direct derivation of neural rosettes from cloned bovine blastocysts: a model of early neurulation events and neural crest specification in vitro
- Source :
- Stem cells (Dayton, Ohio). 24(11)
- Publication Year :
- 2006
-
Abstract
- Embryonic stem cells differentiate into neuroectodermal cells under specific culture conditions. In primates, these cells are organized into rosettes expressing Pax6 and Sox1 and are responsive to inductive signals such as Sonic hedgehog (Shh) and retinoic acid. However, direct derivation of organized neuroectoderm in vitro from preimplantation mammalian embryos has never been reported. Here, we show that bovine inner cell masses from nuclear transfer and fertilized embryos, grown on feeders in serum-free medium, form polarized rosette structures expressing nestin, Pax6, Pax7, Sox1, and Otx2 and exhibiting interkinetic nuclear migration activity and cell junction distribution as in the developing neural tube. After in vitro expansion, neural rosettes give rise to p75-positive neural crest precursor cell lines capable of long-term proliferation and differentiation in autonomic and sensory peripheral neurons, glial cells, melanocytes, smooth muscle cells, and chondrocytes, recapitulating in vitro the unique plasticity of the neural crest lineage. Challenging the rosette dorsal fate by early exposure to Shh induces the expression of ventral markers Isl1, Nkx2.2, and Nkx6.1 and differentiation of mature astrocytes and neurons of central nervous system ventral identity, demonstrating appropriate response to inductive signals. All together, these findings indicate that neural rosettes directly derived from cloned and fertilized bovine embryos represent an in vitro model of early neural specification and differentiation events. Moreover, this study provides a source of highly proliferative neural crest precursor cell lines of wide differentiation potential for cell therapy and tissue engineering applications.
- Subjects :
- Central Nervous System
animal structures
Neuroectodermal Tumors
Mice, SCID
Biology
Mice
Mice, Inbred NOD
Neurosphere
medicine
Animals
Cell Lineage
Hedgehog Proteins
RNA, Messenger
Cell Shape
Cells, Cultured
Embryonic Stem Cells
Cell Proliferation
MSX1 Transcription Factor
Neurons
Neural fold
Neuroectoderm
SOXE Transcription Factors
Neural tube
High Mobility Group Proteins
Neural crest
PAX7 Transcription Factor
Cell Differentiation
Forkhead Transcription Factors
Cell Biology
Cell biology
Clone Cells
Neuroepithelial cell
DNA-Binding Proteins
medicine.anatomical_structure
Neurulation
Homeobox Protein Nkx-2.2
Neural Crest
Astrocytes
Blastocyst Inner Cell Mass
embryonic structures
Immunology
Molecular Medicine
Cattle
Snail Family Transcription Factors
Neural plate
Developmental Biology
Signal Transduction
Stem Cell Transplantation
Transcription Factors
Subjects
Details
- ISSN :
- 10665099
- Volume :
- 24
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- Stem cells (Dayton, Ohio)
- Accession number :
- edsair.doi.dedup.....76eaa70f4578bef2fc82ea633ce3016a