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Sphere forming ability was enhanced by embryo protein treatment. Sphere forming rates were significantly higher in the EP treatment group than in the 11.7‐h or control groups. Manual sphere counts were conducted until day 20 after single cell seeding with each protein treatment. EP, 50% epiboly stage zebrafish embryo extracts. 11.7 h, zebrafish embryo extracts at the segmentation period. Two‐tailed P‐value < 0.0429 and number of pairs = 6 by paired t‐test.
