Regenerative Medicine (2003) / Chapter Skim
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8 Neuroectodermal differentiation from mouse multipotent adult progenitor cells
8 Neuroectodermal differentiation from mouse multipotent adult progenitor cells
Pages 38-44
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From page 38... ...
Abbreviations: BM, bone marrow; NSC, neural stem ceil; MAPC, muitipotent acluit progenitor ceil; m, mouse; ES, embryonic stem; FGF, fibrobiast growth factor; bFGF, basic FGF; SHH, Sonic Hecigehog; BDNF, brain-clerivecl neurotrophic factor; En, embryonic clay n; NF, neurofiiament; MBP, myeiin basic protein; TH, tyrosine hyciroxylase; DDC, clopa-clecarboxylase; TrH, tryptophan hyciroxylase; GABA, y-aminobutyric acicl; GFAP, gliai fibriiiary aciclic protein; TTX, tetroclotoxin; PDGF, piateiet-clerivecl growth factor; eGFP, enhancecl GFP. iTo whom corresponclence shouicl be aciciressecl at: University of Minnesota, MMC 716, 422 Deiaware Street SE, Minneapolis, MN 55455.
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From page 39... ...
GAAGCTCCATATCCCTGGGTGGAAAG CCTCCTCGCGCATGAAGAT CGTCTGTGTGCCTGACACATT AACAGGTCTCCCCGCATCT CACCCTCAGGAACAGAGTGACTT TCTTGACCATCATCTTCTCCAGATC TCTGGAGTTAAGAAATCGGAGCTG GCCTCTGTTCTCCAGCTTGCT GCCGCTCTAGGGACTCGTT ATGCTCTCTGGCTCCTTGGC TGGGCAGGCATGGGC ATGGGCACATTGTGCTTCTG ACACAGCCCAAACTCCACAGT TGACGTTTCTCAGGCATTAAGC GGTGCACTTGCTTGTGCAGT 211 50 141 100 107 102 255 1 1 3 165 153 68 100 100 120 242 antibody and fluorescein or Cy3- or CyS-coupled anti-mouse-, goat-, or rabbit-IgG antibodies. Between each step, slides were washed with PBS plus 1% BSA (Sigma)
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From page 40... ...
-BB and induced to differentiate by the removal of PDGF and addition of bFGF (23) , we initially replated undifferentiated MAPCs at 10,000 cells per cm2 on fibronectin-coated plates or chamberslides, removed epidermal growth factor, PDGF, and leukemia inhibitory factor, and added 100 ng/ml bFGF.
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From page 41... ...
In contrast, after day 5, spiking cells represented between 71% and 100% of cells studied at each time period. Voltage-clamp experiments showed that spiking MAPCs expressed a rapidly inactivating inward current.
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From page 42... ...
Differentiation of marrow pluripotent stem cells to neuron-like cells follows similar developmental steps as described for ES cells and NSCs. Similar to neurons derived from NSCs or ES cells in vitro, in vitro-differentiated MAPCs also exhibit electrophysiological characteristics of neurons.
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From page 43... ...
Rodent or human neuroprogenitors can also be expanded with epidermal growth factor alone, while undergoing initial -differentiation to astrocytes and oligodendrocytes but less neuronal cells, or bFGF alone, while undergoing initial differentiation to neurons and to a lesser extent glia (41~. We demonstrate here that differentiation of mMAPCs to cells with neuroectodermal characteristics occurs by initial culture in the presence of bFGF as the sole cytokine, and that this is associated with the activation of transcription factors known to be important in neural commitment in viva and differentiation from NSCs and mES cells in vitro.
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From page 44... ...
12. Mezey, E., Chandross, K
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