Results: 3

1.
Figure 1

Figure 1. Characterization of the neural derivatives of the VUB03-DM1 cell line.. From: Recurrent genomic instability of chromosome 1q in neural derivatives of human embryonic stem cells.

(A) Morphological features of NSCs at passage 48 derived from VUB03-DM1. (B and C) VUB03-DM1 passage 48 NSCs expressed the neural marker SOX2. Note that cells did not express the neuron-specific ELAV/Hu family members HuC, HuD (HuCD) (B) and TUBB3 (C). (D) Morphological features of neurons derived from VUB03-DM1 passage 48 NSCs. (E and F) Neurons expressing the neuronal markers HuCD (E) and TUBB3 (F) generated after 20 days of differentiation from VUB03-DM1 passage 48 NSCs. Scale bars: 20 μm.

Christine Varela, et al. J Clin Invest. 2012 February 1;122(2):569-574.
2.
Figure 2

Figure 2. Control of genetic stability in the VUB03-DM1 line.. From: Recurrent genomic instability of chromosome 1q in neural derivatives of human embryonic stem cells.

(A) G-banding analysis of the undifferentiated VUB03-DM1 passage 67 (P67) hESCs. (B) mFISH analysis of neural derivatives of VUB03-DM1 passage 34 NSCs showing a segment of chromosome 1 translocated onto chromosome 13p. (C) Partial karyotype showing a segment of chromosome 1 translocated onto chromosomes 5p, 8q, and 13q. (D) Arm-specific chromosome painting revealed 2 copies of chromosome arms 1p (green) and 3 copies of chromosome arm 1q (red). (E) In situ hybridization of a juxtacentromeric-specific probe detecting the 1q12 region showing 3 signals on metaphases (arrows).

Christine Varela, et al. J Clin Invest. 2012 February 1;122(2):569-574.
3.
Figure 3

Figure 3. Differentiation potential in vitro and in vivo.. From: Recurrent genomic instability of chromosome 1q in neural derivatives of human embryonic stem cells.

(AF) In vitro neuronal differentiation potential. 3 weeks after growth factor withdrawal, normal VUB01 passage 18 NSCs (A), normal VUB05-HD passage 15 NSCs (B), and mutant VUB05-HD (batch b) passage 61 NSCs (C) differentiated in vitro into TUBB3-positive neurons. (D) Number of neurons generated with normal VUB01 and VUB05 and mutant VUB05-HD (batch a and b) NSC lines, determined using the neuronal nuclear marker HuCD. Samples of normal and mutant cell lines were differentiated in at least 3 independent experiments. The proportion of HuCD-positive cells (± SEM) was determined in at least 1,000 cells per sample in randomly picked fields. NSC lines bearing 1q duplication failed to differentiate into neurons, except mutant VUB05-HD (batch b). (E and F) 2 weeks after growth factor withdrawal, normal VUB01 passage 18 NSCs (E) differentiated in vitro into neurons, whereas mutant VUB01 passage 71 NSCs (F) failed to give rise to neurons and died. (G and H) In vivo differentiation potential. 7 weeks after grafting, human nestin- and MAP2-positive cells were observed with rat brain transplants of normal VUB01 passage 21 NSCs (G), but not with mutant VUB01 passage 74 NSCs (H). Scale bars: 50 μm.

Christine Varela, et al. J Clin Invest. 2012 February 1;122(2):569-574.

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