GSM4680703: siCTRL-2; Homo sapiens; RNA-Seq
Human dental follicle was obtained from the unerupted third molar with undeveloped root. Patients aged 12-16 years undergoing the third molar extraction in West China Hospital of Stomatology, Sichuan University would meet the inclusion criteria. Patients with history of systemic disease, or undergoing maxillofacial surgery, whole body or partial radiotherapy, chemotherapy, periodontitis, oral mucosal disease or smoking were excluded. All the procedures are approved by Institutional Review Board and the informed consent of patients. After extraction, the dental follicle was instantly immersed into the phosphate buffer saline (PBS, Gibco) with 1× Penicillin-Streptomycin (Liquid, Gibco), i.e. PBS with 100 units/mL penicillin and 100 µg/mL streptomycin. The dental follicle was cut into size of 1mm3 pieces,  and digested with PBS with Type I Collagenase (3mg/ml, Gibco) and Dispase (3 mg/ml, Gibco) for 1 hour in 37℃ water bath with agitation.  The digested tissue suspension was cultured in 21cm2 petri dish in alpha minimum Eagle's medium (α-MEM, HyClone) with 20% fetal bovine serum (FBS, Gibco) and 1× Penicillin-Streptomycin. Incubated at 37℃ with 5% CO2, the culture medium was changed every 2 days. When cell confluence rate reached 80%, hDFCs were passaged and subcultured in α-MEM with 10% FBS and 1× Penicillin-Streptomycin. hDFCs at passage 3 was applied for the subsequent research.  For osteogenic induction, hDFCs were cultured with osteogenic medium supplemented with 50 μM ascorbic acid (Sigma), 10 nM dexamethasone (Sigma) and 10 mM β-glycerophosphate (Sigma). RNA libiaries were prepared for sequencing using standard Illumina protocols.
GEO Accession
GSM4680703