Embryonic mescencephalon derived neurospheres contain progenitors as well as differentiated neurons and glia.
OBJECTIVE
Stem cells and progenitor cells in the central nervous system may have potential for therapeutic use in patients with degenerative diseases or after injury. Neural precursor cells can be grown in culture in the presence of mitogens as aggregates termed neurospheres (NSs), as a source of proliferating progenitor cells. Withdrawal of mitogen and allowing the NSs to adhere to a substrate is the conventional way to study the differentiation potential of the progenitor cells propagated in NSs form. Here we asked if differentiation occurs within NSs cultured in the normal manner, in the presence of mitogen.


METHODS
We used non-passaged NSs derived from E13.5 mouse ventral mesencephalon.


RESULTS
The NSs contained not only progenitor cells but also phenotypically-differentiated neurons and glia, in the presence of mitogen. Extracellular matrix molecules (fibronectin, laminin and collagen type IV) were also detected within these NSs, which may aid in the differentiation of progenitors inside the NSs. The cell types within NSs were also organized in a way that the differentiated cells were found in the inner cell mass while progenitors were found in the outer region. Additionally, the proportion of differentiated cell types within the NSs was also affected by exposure to different mitogens. Moreover, when placed together in to co-culture, dissociated embryonic striatal and mesencephalic cells aggregated spontaneously to form mixed NSs, enhancing the eventual differentiation into dopaminergic neurons from progenitors within these NSs.


CONCLUSIONS
Therefore, the NSs contained progenitor cells and differentiated neurons and glial cells. In addition, NS culture system can be used to study cellular differentiation in vitro in non-adherent conditions.
Khaing
Zin Z
ZZ
The Fishberg Department of Neuroscience, Mount Sinai School of Medicine, New York, NY, USA. zin@che.utexas.edu
Roberts
James L
JL
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D020287:In Situ Nick-End Labeling / Q000379:methods; D008636:Mesencephalon / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009457:Neuroglia / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D011247:Pregnancy; D014404:Tubulin / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D005904:Glial Fibrillary Acidic Protein; C488416:Tubb3 protein, rat; D014404:Tubulin; D005346:Fibroblast Growth Factors; D004815:Epidermal Growth Factor; D014446:Tyrosine 3-Monooxygenase