Antibiotic antimycotic

The umbilical cord sample was collected from mothers undergoing caesarean section in Hospital Canselor Tuanku Muhriz with consent (ethical approval: UKM PPI/111/8/JEP-2023-033). Briefly, arteries and veins were removed from the umbilical cord before mincing into small pieces for enzymatic digestion using 0.6% collagenase type I (Worthington, Lakewood, NJ, USA) for 1 h at 37 °C in a shaking incubator. The isolated cells were cultured in low-glucose Dulbecco’s modified Eagle medium (LG-DMEM; Sigma-Aldrich, St. Louis, MO, USA) with 1% GlutaMAX™ (Gibco, Grand Island, NY, USA), 1% antibiotic–antimycotic (Capricorn, Ebsdorfergrund, Germany), and 10% FBS (Capricorn, Ebsdorfergrund, Germany), 10% H-hPL, or 10% Fd-hPL.

UC-MSCs at passage 4 were seeded in six-well plates with a seeding density of 3000 cells/cm² in LG-DMEM with 1% GlutaMAX™ (Gibco, Grand Island, NY, USA), 1% antibiotic–antimycotic (Capricorn, Ebsdorfergrund, Germany), and 10% FBS (Capricorn, Ebsdorfergrund, Germany), H-hPL, or Fd-hPL. The morphological changes, growth pattern, and confluency of cultured UC-MSCs were observed and captured using an inverted microscope at a magnification of 40×. Five points per well were captured. The length, width, and size of the cells were analyzed using ImageJ software, version 1.53k. Then, the cells were trypsinized with 0.05% trypsin-EDTA (Gibco, Grand Island, NY, USA) when the cell confluency reached 70%–80%. The cells were stained with 0.4% trypan blue solution (Corning^®, Manassas, VA, USA) and the number of viable and non-viable cells was counted using a hemocytometer. The population doubling time (PDT) was calculated using the following formula: $$\mathrm{PDT}\left(\mathrm{hour}\right)=\frac{\mathrm{t}\log 2}{\log \mathrm{N}2-\log \mathrm{N}1}$$
where t denotes time in culture; N2 denotes the cell number at the end of the passage; N1 denotes the cell number seeded at the beginning of the passage.