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URL: https://dx.doi.org/DOI:10.17504/protocols.io.iegcbbw

Authors: yuejun wang

Summary: Teeth were first disinfected with 75% ethanol and then washed with phosphate-buffered saline. Briefly, PDLSCs were separated from periodontal ligament in the middle one-third of the root. Subsequently, MSCs were digested in a solution of 3 mg/mL collagenase type I (Worthington Biochemical Corp., Lakewood, NJ, USA) and 4 mg/mL dispase (Roche Diagnostics Corp., Indianapolis, IN, USA) for 1 h at 37°C. Single-cell suspensions were obtained by cell passage through a 70-μm strainer (Falcon, BD Labware, Franklin Lakes, NJ, USA). MSCs were grown in a humidified, 5% CO2 incubator at 37°C in DMEM alpha modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA), supplemented with 15% fetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA), 2 mmol/L glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin (Invitrogen, Carlsbad, CA, USA). The culture medium was changed every 3 days. MSCs at passages 3-5 were used in subsequent experiments. Human embryonic kidney 293T cells were maintained in complete DMEM with 10% FBS, 100 U/mL penicillin and 100 µg/mL streptomycin.

Associated Publications: Wang Y, Liu Y, Fan Z, Liu D, Wang F, Zhou Y (2017) IGFBP2 enhances adipogenic differentiation potentials of mesenchymal stem cells from Wharton's jelly of the umbilical cord via JNK and Akt signaling pathways. PLoS ONE 12(8): e0184182. doi: 10.1371/journal.pone.0184182

Affiliations: Department of Prosthodontics, Peking University School and Hospital of Stomatology

External URL: https://doi.org/10.1371/journal.pone.0184182

Version: 1

Publication Date: 2017

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