First, HUVECs were seeded into 24-well plates at a concentration of 1.0 × 104 cells/well. The cells were cultured overnight in a standard culture medium supplemented with 10% FBS, the medium was discarded, and 1 ml of the conditioned medium from the different experimental groups supplemented with 5% FBS (only DMEM was used as the control) was added. At different time points (placental cell-derived conditioned medium groups: 1, 2, 3, to 6 days; sub-cultured placental tissue-derived conditioned medium groups: 12, 24, 36, and 48 h), the cells were harvested and counted. The proliferation ability of HUVECs was measured by counting the number of cells using a Countstar automated cell counter (ALIT Life Science, USA).

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