Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Oct 15;90(20):9683–9687. doi: 10.1073/pnas.90.20.9683

Reconstruction of neural tube-like structures in vitro from primary neural precursor cells.

Y Tomooka 1, H Kitani 1, N Jing 1, M Matsushima 1, T Sakakura 1
PMCID: PMC47634  PMID: 8415762

Abstract

Vertebrate central nervous system develops from a neural tube derived from the embryonic ectoderm. In mouse, the neural tube around embryonic day 10 primarily consists of neural precursor cells (NPCs). During the development of embryonic central nervous system, NPCs proliferate and migrate outward; thus later stages show NPCs toward the lumen of the neural tube and neurofilament-positive differentiated cells toward the periphery. In conventional liquid culture, NPCs isolated from mouse on embryonic day 10 proliferate and differentiate into neurofilament-positive neurons. In the present communication, we show that fragments of neural tubes and aggregates of NPCs, when placed into collagen gel matrix, form three-dimensional structures which resemble the neural tube formed in vivo in the developing embryos. Even dissociated NPCs form the three-dimensional structures in the collagen gel matrix. Our results indicate that individual NPCs or fragments of neural tubes carry morphogenetic information which allows them to reconstruct neural tube-like structures in vitro.

Full text

PDF
9683

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brun R. B., Garson J. A. Neurulation in the Mexican salamander (Ambystoma mexicanum): a drug study and cell shape analysis of the epidermis and the neural plate. J Embryol Exp Morphol. 1983 Apr;74:275–295. [PubMed] [Google Scholar]
  2. Chan W. Y., Tam P. P. The histogenetic potential of neural plate cells of early-somite-stage mouse embryos. J Embryol Exp Morphol. 1986 Jul;96:183–193. [PubMed] [Google Scholar]
  3. Cochard P., Paulin D. Initial expression of neurofilaments and vimentin in the central and peripheral nervous system of the mouse embryo in vivo. J Neurosci. 1984 Aug;4(8):2080–2094. doi: 10.1523/JNEUROSCI.04-08-02080.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DeLong G. R. Histogenesis of fetal mouse isocortex and hippocampus in reaggregating cell cultures. Dev Biol. 1970 Aug;22(4):563–583. doi: 10.1016/0012-1606(70)90169-7. [DOI] [PubMed] [Google Scholar]
  5. Easter S. S., Jr, Ross L. S., Frankfurter A. Initial tract formation in the mouse brain. J Neurosci. 1993 Jan;13(1):285–299. doi: 10.1523/JNEUROSCI.13-01-00285.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FUJITA S. The matrix cell and cytogenesis in the developing central nervous system. J Comp Neurol. 1963 Feb;120:37–42. doi: 10.1002/cne.901200104. [DOI] [PubMed] [Google Scholar]
  7. Iguchi T., Uchima F. D., Ostrander P. L., Bern H. A. Growth of normal mouse vaginal epithelial cells in and on collagen gels. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3743–3747. doi: 10.1073/pnas.80.12.3743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imagawa W., Tomooka Y., Nandi S. Serum-free growth of normal and tumor mouse mammary epithelial cells in primary culture. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4074–4077. doi: 10.1073/pnas.79.13.4074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kitani H., Shiurba R., Sakakura T., Tomooka Y. Isolation and characterization of mouse neural precursor cells in primary culture. In Vitro Cell Dev Biol. 1991 Aug;27A(8):615–624. doi: 10.1007/BF02631104. [DOI] [PubMed] [Google Scholar]
  10. Lendahl U., Zimmerman L. B., McKay R. D. CNS stem cells express a new class of intermediate filament protein. Cell. 1990 Feb 23;60(4):585–595. doi: 10.1016/0092-8674(90)90662-x. [DOI] [PubMed] [Google Scholar]
  11. Martins-Green M., Erickson C. A. Development of neural tube basal lamina during neurulation and neural crest cell emigration in the trunk of the mouse embryo. J Embryol Exp Morphol. 1986 Nov;98:219–236. [PubMed] [Google Scholar]
  12. Matsuda M. Serum proteins enhance aggregate formation of dissociated fetal rat brain cells in an aggregating culture. In Vitro Cell Dev Biol. 1988 Oct;24(10):1031–1036. doi: 10.1007/BF02620877. [DOI] [PubMed] [Google Scholar]
  13. Montesano R., Schaller G., Orci L. Induction of epithelial tubular morphogenesis in vitro by fibroblast-derived soluble factors. Cell. 1991 Aug 23;66(4):697–711. doi: 10.1016/0092-8674(91)90115-f. [DOI] [PubMed] [Google Scholar]
  14. Moury J. D., Jacobson A. G. Neural fold formation at newly created boundaries between neural plate and epidermis in the axolotl. Dev Biol. 1989 May;133(1):44–57. doi: 10.1016/0012-1606(89)90295-9. [DOI] [PubMed] [Google Scholar]
  15. Rutishauser U. Developmental biology of a neural cell adhesion molecule. Nature. 1984 Aug 16;310(5978):549–554. doi: 10.1038/310549a0. [DOI] [PubMed] [Google Scholar]
  16. Schoenwolf G. C. Microsurgical analyses of avian neurulation: separation of medial and lateral tissues. J Comp Neurol. 1988 Oct 22;276(4):498–507. doi: 10.1002/cne.902760404. [DOI] [PubMed] [Google Scholar]
  17. Schoenwolf G. C., Smith J. L. Mechanisms of neurulation: traditional viewpoint and recent advances. Development. 1990 Jun;109(2):243–270. doi: 10.1242/dev.109.2.243. [DOI] [PubMed] [Google Scholar]
  18. Sheffield J. B., Moscona A. A. Electron microscopic analysis of aggregation of embryonic cells: the structure and differentiation of aggregates of neural retina cells. Dev Biol. 1970 Sep;23(1):36–61. doi: 10.1016/s0012-1606(70)80006-9. [DOI] [PubMed] [Google Scholar]
  19. Smedley M. J., Stanisstreet M. Calcium and neurulation in mammalian embryos. J Embryol Exp Morphol. 1985 Oct;89:1–14. [PubMed] [Google Scholar]
  20. Sternberg J., Kimber S. J. Distribution of fibronectin, laminin and entactin in the environment of migrating neural crest cells in early mouse embryos. J Embryol Exp Morphol. 1986 Feb;91:267–282. [PubMed] [Google Scholar]
  21. Takeichi M. Cadherin cell adhesion receptors as a morphogenetic regulator. Science. 1991 Mar 22;251(5000):1451–1455. doi: 10.1126/science.2006419. [DOI] [PubMed] [Google Scholar]
  22. Tomooka Y., Harris S. E., McLachlan J. A. Growth of seminal vesicle epithelial cells in serum-free collagen gel culture. In Vitro Cell Dev Biol. 1985 Apr;21(4):237–244. doi: 10.1007/BF02620935. [DOI] [PubMed] [Google Scholar]
  23. Yang J., Larson L., Nandi S. Three-dimensional growth and morphogenesis of mouse submandibular epithelial cells in serum-free primary culture. Exp Cell Res. 1982 Feb;137(2):481–485. doi: 10.1016/0014-4827(82)90057-x. [DOI] [PubMed] [Google Scholar]
  24. van Straaten H. W., Hekking J. W., Beursgens J. P., Terwindt-Rouwenhorst E., Drukker J. Effect of the notochord on proliferation and differentiation in the neural tube of the chick embryo. Development. 1989 Dec;107(4):793–803. doi: 10.1242/dev.107.4.793. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES