Liberase mnp s enzyme

Normal human third molar teeth indicated for extraction were collected from patients aged 20–28 years at the Aichi-Gakuin University Dental Hospital, in accordance with the approved guidelines set by the School of Dentistry, Aichi-Gakuin University, and the National Center for Geriatrics and Gerontology Research Institute. All experimental protocols were approved by the National Center for Geriatrics and Gerontology Research Institute. Informed consent was obtained from all subjects involved in this experiment and donor information for used DPSCs can be found as Supplementary Table S2. The pulp was gently removed using a sterile dental probe and the collected pulp tissue was dissected and digested in 0.2% Liberase MNP-S enzyme (Roche, Germany). The isolated dental pulp cells (DPCs) were cultured in Dulbecco modified eagle’s medium (DMEM) supplemented with 10% human serum collected from healthy consenting adult donors and Antibiotic-Antimitotic solution (life technologies) containing; 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 0.25 μg/mL of Fungizone®, final concentration. Cells were selected on the basis of their ability to adhere to the dish; non-adherent cells were removed during medium replacement after 4–5 days in culture. DPSCs from up to 4 donors were used to conduct the various assays.

Normal human third molar teeth indicated for extraction were collected from patients aged 20–28 years at the Aichi-Gakuin University Dental Hospital, in accordance with the approved guidelines set by the School of Dentistry, Aichi-Gakuin University, and the National Centre for Geriatrics and Gerontology Research Institute. All experimental protocols were approved by the National Centre for Geriatrics and Gerontology Research Institute. Informed consent was obtained from all subjects involved in this experiment and donor information for used dental pulp derived mesenchymal stem cells can be found in Supplementary Table S1 in Supplementary Material available online at http://dx.doi.org/10.1155/2016/8102478. The pulp was gently removed using a sterile dental probe and the collected pulp tissue was dissected and digested in 0.2% Liberase MNP-S enzyme (Roche, Germany). The isolated dental pulp cells were cultured in Dulbecco Modified Eagle's Medium (DMEM) supplemented with 10% human serum and Antibiotic-Antimitotic solution (Gibco, Life Technologies) containing 500 units/mL of penicillin, 500 μg/mL of streptomycin, and 1.25 μg/mL of Fungizone^®. Cells were selected on the basis of their ability to adhere to the dish; nonadherent cells were removed during medium replacement after 4-5 days in culture. Subsequent experiments have been performed in triplets on 3 different samples.