This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kawata, M. Articles by Takamizawa, H. Search for Related Content PubMed PubMed Citation Articles by Kawata, M. Articles by Takamizawa, H.

Neural Rosette Formation within in Vitro Spheroids of a Clonal Human Teratocarcinoma Cell Line, PA-1/NR: Role of Extracellular Matrix Components in the Morphogenesis¹

Departments of Obstetrics and Gynecology [M. K., S. S., H. K., H. T.] and Anatomy [K. K.], School of Medicine, Chiba University, 1-8-1 Inohana, Chiba City, Chiba 280, Japan

From the human teratocarcinoma-derived cell line PA-1, we established a clonal line, PA-1/NR, that stably produced a distinct cellular arrangement of neural rosettes when cultured as in vitro multicellular spheroids for 3 weeks. On immunofluorescence staining and fluorescence-activated cell sorter analyses, PA-1/NR cells in monolayer expressed the neuroectoderm-associated antigens HNK-1, NC-1, and A2B5 and the neuroblastoma-associated antigens KP-NAC8 and KP-NAC10 but lacked human embryonal carcinoma antigens, SSEA-3 or K21 antigen. Here, we investigated the developmental process of rosette formation with respect to morphological features, distribution of mitotic cells, and expression of multiple lineage-related markers and extracellular matrix (ECM) components.

Ultrastructural examination of these rosettes disclosed a well-defined cavity radially surrounded by wedge-shaped or pseudostratified cells, apical microvilli and junctional complexes, and basal laminae and collagen fibrils at their basal surface. In these rosettes, many proliferating cells were detected by the immunohistochemical staining of cells incorporating bromodeoxyuridine. PA-1/NR spheroids consistently displayed neuron-specific enolase, S-100 protein, and vimentin but not glial fibrillary acidic protein, neurofilament proteins, or myelin basic protein.

The rosette formation accompanied a strikingly polarized and over-lapped deposition of ECM components including tenascin-carrying HNK-1 epitopes, laminin, type IV collagen, heparan, and chondroitin sulfate proteoglycans. Immunoblotting analyses showed that laminin B1 and B2 chains were constitutively expressed, whereas a fully assembled form of laminin and type IV collagen appeared only after spheroid development, suggesting that these ECM components play a morphogenetically important role in rosette formation. Close similarities between these rosettes and the neural tube of humans and experimental animals in the morphogenetic process and ECM formation lead us to propose that the PA-1/NR spheroids provide an in vitro model for the study of the earliest stage of human neurogenesis.

¹ This work was supported by grants from the Ministry of Education, Science and Culture, Japan.

² To whom reprint requests should be addressed.

Received 11/19/90. Accepted 3/ 8/91.

This article has been cited by other articles:

				M. Denham, B. J. Conley, F. Olsson, L. Gulluyan, T. J. Cole, and R. Mollard A murine respiratory-inducing niche displays variable efficiency across human and mouse embryonic stem cell species Am J Physiol Lung Cell Mol Physiol, May 1, 2007; 292(5): L1241 - L1247. [Abstract] [Full Text] [PDF]