Robust generation of oligodendrocyte progenitors from human neural stem cells and engraftment in experimental demyelination models in mice.
BACKGROUND
Cell-based therapy holds great promises for demyelinating diseases. Human-derived fetal and adult oligodendrocyte progenitors (OPC) gave encouraging results in experimental models of dysmyelination but their limited proliferation in vitro and their potential immunogenicity might restrict their use in clinical applications. Virtually unlimited numbers of oligodendroglial cells could be generated from long-term self-renewing human (h)-derived neural stem cells (hNSC). However, robust oligodendrocyte production from hNSC has not been reported so far, indicating the need for improved understanding of the molecular and environmental signals controlling hNSC progression through the oligodendroglial lineage. The aim of this work was to obtain enriched and renewable cultures of hNSC-derived oligodendroglial cells by means of epigenetic manipulation.


RESULTS
We report here the generation of large numbers of hNSC-derived oligodendroglial cells by concurrent/sequential in vitro exposure to combinations of growth factors (FGF2, PDGF-AA), neurotrophins (NT3) and hormones (T3). In particular, the combination FGF2+NT3+PDGF-AA resulted in the maintenance and enrichment of an oligodendroglial cell population displaying immature phenotype (i.e., proliferation capacity and expression of PDGFRalpha, Olig1 and Sox10), limited self-renewal and increased migratory activity in vitro. These cells generate large numbers of oligodendroglial progeny at the early stages of maturation, both in vitro and after transplantation in models of CNS demyelination.


CONCLUSIONS
We describe a reliable method to generate large numbers of oligodendrocytes from a renewable source of somatic, non-immortalized NSC from the human foetal brain. We also provide insights on the mechanisms underlying the pro-oligodendrogenic effect of the treatments in vitro and discuss potential issues responsible for the limited myelinating capacity shown by hNSC-derived oligodendrocytes in vivo.
Neri
Margherita
M
San Raffaele Scientific Institute, San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milano, Italy.
Maderna
Claudio
C
Ferrari
Daniela
D
Cavazzin
Chiara
C
Vescovi
Angelo L
AL
Gritti
Angela
A
D000818:Animals; D018929:Cell Culture Techniques; D003711:Demyelinating Diseases / Q000628:therapy*; D006085:Graft Survival; D006801:Humans; D051379:Mice; D023421:Models, Animal; D039902:Multipotent Stem Cells / Q000166:cytology; D009474:Neurons / Q000166:cytology; D009836:Oligodendroglia / Q000166:cytology / Q000637:transplantation*; D033581:Stem Cell Transplantation / Q000379:methods*; D014183:Transplantation, Heterologous
Nat Biotechnol. 1998 Nov;16(11):1033-9
9831031
PLoS One. 2008 Oct 02;3(10):e3310
18836552
Glia. 2006 Jan 15;53(2):167-81
16206164
Stem Cells. 2007 Sep;25(9):2312-21
17556596
Cell Stem Cell. 2007 Jun 7;1(1):39-49
18371333
J Neurosci Res. 1999 Aug 1;57(3):304-14
10412021
Curr Biol. 2003 Jan 21;13(2):151-5
12546790
Neurochem Res. 2007 Apr-May;32(4-5):783-97
17186374
J Cell Biol. 2004 Jan 5;164(1):111-22
14709544
Neurosci Lett. 2009 Jun 12;456(3):112-9
19429145
J Neurosci Res. 2009 Nov 15;87(15):3438-46
19739249
Exp Cell Res. 2006 Jul 1;312(11):2107-20
16631163
Nat Neurosci. 2008 Dec;11(12):1392-401
18849983
Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14069-74
16172374
Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14720-5
11121071
Curr Opin Neurol. 2008 Jun;21(3):278-83
18451710
J Neurosci. 1999 Nov 15;19(22):9986-95
10559406
J Neurosci Res. 2002 Sep 15;69(6):976-86
12205691
PLoS One. 2007 Aug 01;2(8):e689
17668067
Gene Ther. 2006 Mar;13(5):379-88
16355116
J Neurosci Res. 1992 Aug;32(4):494-506
1356158
Int J Dev Neurosci. 2009 Dec;27(8):769-78
19720126
Nature. 1994 Jan 27;367(6461):371-5
8114937
J Neurosci Res. 2002 Sep 15;69(6):966-75
12205690
J Neurobiol. 1999 Sep 15;40(4):497-512
10453052
Glia. 2004 Sep;47(4):314-24
15293229
Genes Dev. 2002 Jan 15;16(2):165-70
11799060
Proc Natl Acad Sci U S A. 2006 May 16;103(20):7853-8
16682644
J Neurosci. 2009 Jun 17;29(24):7649-57
19535576
Nature. 1988 Jun 9;333(6173):560-2
3287176
Mol Cell Neurosci. 2000 Oct;16(4):422-39
11085879
Nat Med. 2004 Jan;10(1):93-7
14702638
Proc Natl Acad Sci U S A. 2004 Aug 10;101(32):11839-44
15280535
Stem Cells Dev. 2009 Sep;18(7):1043-58
19327007
Cell. 1992 Jul 10;70(1):31-46
1623522
Neuroscience. 2007 Aug 24;148(2):535-40
17681700
J Comp Neurol. 2004 Jul 19;475(2):211-9
15211462
Neuropathol Appl Neurobiol. 1996 Jun;22(3):199-206
8804021
Cell Stem Cell. 2008 Jun 5;2(6):553-65
18522848
Glia. 2005 May;50(3):212-22
15712210
Glia. 2003 Nov;44(2):153-65
14515331
Exp Neurol. 1999 Aug;158(2):265-78
10415135
Mol Cell Neurosci. 1996 Aug;8(2/3):146-56
8954629
Dev Neurosci. 2001;23(4-5):327-37
11756748
Nat Biotechnol. 1998 Nov;16(11):1040-4
9831032
Ann Neurol. 2009 Sep;66(3):343-54
19798728
Clin Pharmacol Ther. 2007 Oct;82(4):453-60
17713467
J Neurosci. 1992 Apr;12(4):1538-47
1556607
J Neurosci. 2008 Oct 8;28(41):10434-42
18842903
Nat Cell Biol. 2002 Nov;4(11):833-41
12379866
Curr Opin Neurol. 2007 Dec;20(6):688-92
17992090
EMBO J. 2002 Dec 2;21(23):6452-60
12456652
Stem Cells. 2009 Jan;27(1):116-25
18403757
J Comp Neurol. 2009 Nov 20;517(3):333-49
19760739
PLoS One. 2007 Jan 17;2(1):e156
17225860
Exp Neurol. 1999 Mar;156(1):71-83
10192778
J Neurosci. 2004 Apr 14;24(15):3816-25
15084663
Neuron. 1998 May;20(5):869-82
9620692
J Neurosci. 2002 Jan 15;22(2):437-45
11784788
Brain Pathol. 1999 Jul;9(3):499-513
10416990
Development. 2003 Dec;130(26):6599-609
14660548
J Neurotrauma. 1999 Aug;16(8):689-93
10511241
J Neurosci. 1999 Jul 15;19(14):5990-6005
10407037
Nat Biotechnol. 2001 May;19(5):475-9
11329020
Nat Med. 2003 Apr;9(4):439-47
12627226
Glia. 2002 Oct;40(1):65-77
12237844
Development. 2008 Feb;135(4):637-46
18184726
Glia. 2006 Jul;54(1):35-46
16673374
Mol Cell Neurosci. 2008 Jun;38(2):245-58
18450476
J Neurosci Res. 1997 Oct 15;50(2):146-56
9373025
J Neurosci Res. 2000 Feb 1;59(3):421-9
10679779
Rejuvenation Res. 2005 Fall;8(3):131-4
16144466
Nat Neurosci. 2002 Dec;5(12):1271-8
12426573