Differentiation of human adult skin-derived neuronal precursors into mature neurons.
The isolation of autologous neuronal precursors from skin-derived precursor cells extracted from adult human skin would be a very efficient source of neurons for the treatment of various neurodegenerative diseases. The purpose of this study was to demonstrate that these neuronal precursors were able to differentiate into mature neurons. We isolated neuronal precursors from breast skin and expanded them in vitro for over ten passages. We showed that 48% of these cells were proliferating after the first passage, while this growth rate decreased after the second passage. We demonstrated that 70% of these cells were nestin-positive after the third passage, while only 17% were neurofilament M-positive after 7 days of differentiation. These neuronal precursors expressed betaIII tubulin, the dendritic marker MAP2 and the presynaptic marker synaptophysin after 7 days of in vitro maturation. They also expressed the postsynaptic marker PSD95 and the late neuronal markers NeuN and neurofilament H after 21 days of differentiation, demonstrating they became terminally differentiated neurons. These markers were still expressed after 50 days of culture. The generation of autologous neurons from an accessible adult human source opens many potential therapeutic applications and has a great potential for the development of experimental studies on normal human neurons.
Gingras
Marie
M
Laboratoire d'Organogénèse Expérimentale (LOEX), Centre Hospitalier Affilié Universitaire de Québec, Hôpital du St-Sacrement and Département de Chirurgie, Faculté de Médecine, Université Laval, Québec, Canada.
Champigny
Marie-France
MF
Berthod
François
F
D000328:Adult; D000368:Aged; D000943:Antigens, Differentiation / Q000378:metabolism; D034961:Antigens, Nuclear / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage / Q000502:physiology*; D002478:Cells, Cultured; D000076122:Disks Large Homolog 4 Protein; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D012867:Skin / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D016708:Synaptophysin / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
D000943:Antigens, Differentiation; D034961:Antigens, Nuclear; C501444:DLG4 protein, human; D000076122:Disks Large Homolog 4 Protein; D007381:Intermediate Filament Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D016900:Neurofilament Proteins; D016708:Synaptophysin; D014404:Tubulin; C506808:neuronal nuclear antigen NeuN, human; C051774:neurofilament protein H; C061195:neurofilament protein M