Continuous observation of multipotential retinal progenitor cells in clonal density culture.
All neural cell types in the vertebrate retina, except astrocytes, have been shown to develop from multipotential progenitor cells. It is not known, however, to what extent the progenitor cells are heterogeneous in their developmental potential or to what extent cell-cell interactions versus cell-autonomous factors influence the types of cells they become. To address these issues we developed a clonal-density cell culture system where mouse retinal progenitor cells can survive, divide, and differentiate. We followed the development of clones both by continuous time-lapse video microscopy and by daily microscopic observation. We show that even when cultured at clonal density in a homogeneous general environment, where they cannot contact cells outside their own clone, the retinal progenitor cells vary in proliferative capacity, cell cycle time, and in the cell types that they generate. In addition, we show that under these conditions single progenitor cells can generate both neurons and glia, in which case the neurons almost always develop before glial cells, as is the case in vivo.
Jensen
A M
AM
MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, United Kingdom. ajensen@uoneuro.uoregon.edu
Raff
M C
MC
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002470:Cell Survival; D002999:Clone Cells; D005260:Female; D019254:Genes, bcl-2; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018715:Microscopy, Video; D010751:Phosphopyruvate Hydratase / Q000235:genetics; D011247:Pregnancy; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012160:Retina / Q000166:cytology / Q000196:embryology*; D013234:Stem Cells / Q000166:cytology*; D014714:Vertebrates
D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; D010751:Phosphopyruvate Hydratase