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1.
Figure 5

Figure 5. Cell apoptosis analysis of mES cells cultured under SMG or 1G.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

(A) Flow cytometric analysis of cells to assess apoptosis using Annexin V labeling. Experiments were performed thrice and representative analyzes are shown. (B) Quantitative comparison of apoptosis between the 1G Group and the SMG Group. Three apoptotic assays shown in (A) were performed for comparison. Student's t test, *p<0.05.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.
2.
Figure 2

Figure 2. Cell Number Expansion analysis of mES cells after five days incubation under SMG or 1G.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

The initial cell seeding number was 1×104. (A) Cell number determined by counting cells at indicated time points with a standard hemocytometer. (B) The doubling generation curve generated by dividing the cell number with 104 and then transferring the quotient to the logarithm to the base 2. The data represents mean ± SD of three independent experiments. Student's t test, *p<0.05.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.
3.
Figure 1

Figure 1. Differentiation state of mES cells after incubation under SMG or 1G.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

(A) Upper: Representative microphotographs of mES cells of group 1G and group SMG after ALP staining on Day 2 and Day 7, taken at 100× and 200× magnification respectively. Lower: Percentages of ALP-positive colonies on Day 2 and Day 7. More than 50 colonies of each group were checked for the calculation of the positive colony ratio. (B) Evaluation of the relative expression level of Oct4 and Nanog by quantitative real-time PCR on Day 2 and Day 7. Oct4 and Nanog are extensively used pluripotency markers of stem cells. The data represents mean ± SD of three independent experiments.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.
4.
Figure 6

Figure 6. Comet assay analysis of DNA damage and DNA damage repair under SMG or 1G.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

(A) Evaluation of DNA damage by the alkaline comet assay in mES cells after two days incubation under SMG or 1G. (B) Kinetics of comet tail moment in mES cells irradiated with 8 Gy of gamma-ray and incubated in 1G or in SMG evaluated by neutral comet assay. Time points were 0 hr, 1 hr, 2 hr. On the right representative comet assay results were shown, and on the left quantitative comparison of comet tail moments between the 1G group and the SMG group. At least 50 cells of each time point was scored for comet tail moment. The data represents mean ± SD of three independent experiments. Student's t test, *p<0.05.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.
5.
Figure 3

Figure 3. Cell proliferation analyzed by three independent methods from three different aspects.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

Mouse ES cells were cultured under SMG or 1G for indicated time points. Then cells were processed properly for the following analysis: (A) Quantitative comparison of cell cycle distribution between the 1G group and SMG group. Cell cycle distribution was determined by flow cytometry at 4 hr, 6 hr, 12 hr, 24 hr, 48 hr and 72 hr. The data represents mean ± SD of three independent experiments. (B) Evaluation of relative expression levels of PCNA by quantitative real-time PCR on Day 2. The data represents mean ± SD of three independent experiments. (C) Flow cytometry analysis of cells stained with both PI and BrdU after 1day and 2days incubation. BrdU-positive S phase cells were gated. Experiments were performed thrice and representative analyzes are shown.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.
6.
Figure 4

Figure 4. Microgravity induced decrease in adherent mES cells at early exposure hours.. From: Effects of Simulated Microgravity on Embryonic Stem Cells.

Mouse ES cells were cultured in conventional culture condition under SMG or 1G. At indicated time points, adherent cells and detached cells were collected separately. The adhesion rate is the ratio of adhesive cells to the sum of adherent cells and detached cells. (A) The adhesion rate at 2 hr, 4 hr, 6 hr, 8 hr, 10 hr, 12 hr, 24 hr and 48 hr. (B) Prolonged exposure time to MG and the enhanced adhesion rate. Cells in Group 1 d+6 hr were cultured for 1 day and then the detached and dead cells were discarded by replacing the culture media with new ones. After that, cells were incubated for another 6 hours. The data represents mean ± SD of three independent experiments. Student's t test, *p<0.05.

Yulan Wang, et al. PLoS One. 2011;6(12):e29214.

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