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. 2021 May 8;44(4):560–566. doi: 10.1016/j.htct.2021.02.005

Table 1.

Details of the five protocols used to culture murine BM-MSCs.

Animal age (weeks) Harvesting Initial cell density Plate Culture medium Serum Medium replacement
Protocol 1 6 Maceration ∼8 × 107cells 25cm2 flask DMEM 10% and 20% FBS 7 days
Protocol 2 6 to 8 Flush 1 × 107cells 25cm2 flask RPMI 2µM L-glutamine 1 × 10−6mol/L hydrocortisone 10% and 20% HS 7 days
Protocol 3 6 to 8 Flush 2 × 107cells 25cm2 flask RPMI 2µM L-glutamine 1 × 10−6mol/L hydrocortisone 20% HS 7 days
Protocol 4 16 Flush 1.75 × 107cells 6-well plate DMEM 10% FBS 72 hours
Protocol 5 8 to 10 Flush and centrifugation ∼8 × 107cells 175cm2 flask RPMI - IMDM 12µM L-glutamine 9% FBS 9% HS 24 hours

DMEM: Dulbecco's Modified Eagle Medium; RPMI-1640: Roswell Park Memorial Institute; IMDM: Iscove's modified Dulbecco's Medium; FBS: Fetal Bovine Serum; HS: horse serum.