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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Mesenchymal stem cell-derived conditioned medium protects vascular grafts of brain-dead rats against in vitro ischemia/reperfusion injury

Fig. 1

A simplified schematic of the bone marrow-derived mesenchymal stem cell (MSC)-conditioned medium (CM) collection protocol. Rats were euthanized with an overdose of pentobarbital sodium (100 mg/kg, intraperitoneally). The bone marrow was isolated by flushing the femurs and tibias with Dulbecco’s phosphate-buffered saline (DPBS). The cells were suspended in MSC Expansion Medium and then incubated at 37 °C with 5% CO2 on cell culture flasks. Primary cells were subcultured 1:3 when 80% confluency was reached. After MSCs reached greater than 80% confluency at passage 3, the medium was aspirated, and MSCs were rinsed 3 times with DPBS. Then, Dulbecco’s modified Eagle’s medium (D-MEM) was added to culture dishes with MSCs, and the culture dishes were put into an incubator for 24 h. Primary CM was collected and centrifuged by ultrafiltration units (4500g for 4 h at 4 °C) to yield concentrated CM

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