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. 2024 Jan 18;11:1293068. doi: 10.3389/fcell.2023.1293068

FIGURE 6.

FIGURE 6

Detrimental effects of the medium of high cell density culture on MEFS at 3.5% O2 (A) Transmitted light images representative of the effects of medium harvested after 5 days of high density cell culture at 3.5% O2 and used as conditioned medium on new SSCs culture in 21% O2 condition, scale bar: 100 μm. (B) Cell number after 7-days culture on MEFS feeder in presence of medium conditioned during 5 days at various cell density (0 and 30,000 cells) and tension of O2 (21% and 3.5% O2), 21% new medium (n = 7), 21% 5 days without cells (n = 6), 21% 5 days high cell density (n = 8), 3.5% 5 days without cells (n = 8), 3.5% 5 days high cell density (n = 6). (C) Cell death in inactivated MEFs feeders grown under 21% and 3.5% O2 culture conditions (n = 3). (D) pO2, pCO2, pH and lactate dosage in media after 4-days culture on MEFs feeder at various SSCs density (0, 1000 and 30,000 cells) and tension of O2 (21% and 3.5% O2), in media on MEFs feeder only (MEFs), and in media without MEFs and SSCs (medium). pO2: 1000 cells, 21% and 3.5% O2, n = 7; 30,000 cells, 21% and 3.5% O2, n = 10; MEFs, 21% and 3.5% O2; n = 3; medium, 21% (n = 3) and 3.5% (n = 4) O2. pCO2: 1000 cells, 21% and 3.5% O2, n = 10; 30,000, 21% (n = 13) and 3.5% (n = 11) O2; MEFs, 21% and 3.5% O2; n = 6; medium, 21% (n = 6) and 3.5% (n = 7) O2. pH: 1000 cells, 21% (n = 13) and 3.5% (n = 7) O2,; 30,000 cells, 21% (n = 16) and 3.5% (n = 10) O2; MEFs, 21% and 3.5% O2; n = 6; medium, 21% (n = 7) and 3.5% O2 (n = 6). Lactate: 1000 cells, 21% and 3.5% O2 (n = 10); 30,000 cells, 21% (n = 10) and 3.5% (n = 8) O2; MEFS, 21% and 3.5% O2; n = 6; medium, 21% and 3.5% O2 (n = 6). (E) Number of cells generated after 7 days cell culture at high cell density on various MEFs feeder cell density in 21% and 3.5% O2 conditions (n = 2). (F) pH of the medium when CO2 tensions of 1% and 3.5% are applied to the culture.