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. 2016 Aug 9;7(2):177–191. doi: 10.1016/j.stemcr.2016.07.009

Figure 1.

Figure 1

KOSR and 2i Culture Enhances ESC Single-Cell Potency

ESCs were derived (D) in and cultured (C) under various conditions; D in serum and C in serum, D in serum and C in 2i, D in 2i and C in 2i, and D in KOSR and C in KOSR.

(A and B) Multiple (5) ESCs were injected into morulae and their contribution to full-term chimeras assessed according to coat color. (A) Graph showing the percentage of chimeras born out of the total number of pups and quantification of coat color chimerism. n Values are provided above bars and refer to the number of independent ESC clones tested for each condition. Error bars indicate SEM. Results for 2i conditions are a combination of ESCs cultured with and without mouse embryonic fibroblasts (MEFs) to control for the effect that MEFs may have in serum and KOSR cultures. No difference was observed with or without MEFs. ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001, Student's unpaired t test. (B) Representative images of litters of mice. Asterisks indicate mice with ESC contribution.

(C and D) ESCs were cultured in serum, 2i, or KOSR and stained for alkaline phosphatase activity. (C) Representative images of ESC colonies. (D) Quantification of ESC colony categories as defined in Figure S1C. Error bars represent SEM of three biological replicates using independent ESC lines. ∗∗∗∗p ≤ 0.0001, two-tailed chi-square test.

(E and F) ESCs were cultured in serum, 2i, or KOSR and then differentiated by removal of LIF for 5 days. Cells were stained for alkaline phosphatase activity. (E) Representative images of differentiated colonies. (F) Quantification of ESC colony categories as defined in Figure S1C. Error bars represent SEM of three biological replicates of independent ESC lines. ∗∗∗∗p ≤ 0.0001, two-tailed chi-square test.

(G–I) Single ESCs were injected into morulae and their contribution to full-term chimeras assessed according to coat color. (G) Graph showing the percentage of chimeras born out of the total number of pups and quantification of coat color chimerism. n Values are provided above bars and refer to the number of independent ESC clones tested for each condition. Error bars indicate SEM. Results for 2i conditions are a combination of ESCs cultured with and without MEFs to control for the effect that MEFs may have in serum and KOSR cultures. No difference was observed with or without MEFs. (H and I) Representative images of litters of mice. Asterisks indicate mice with ESC contribution.

See also Figure S1.