Methods : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
The isolation and characterization of DFCs and dNC-PCs were described in previous studies [4,7,12]. DFCs were routinely cultivated in DMEM (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 100 μg/ml penicillin/streptomycin (standard cell culture medium). dNC-PCs were cultivated in DMEM (Sigma-Aldrich) supplemented with 15% fetal bovine serum (Sigma-Aldrich) and 100 μg/ml penicillin/streptomycin (standard cell culture medium). For experiments, both cell types were used after passage 6. DFCs and dNC-PCs expressed typical markers for dental stem cells such as CD105, Nestin, and STRO-1 (Additional file 1: Figure S1).
Five milliliter of PA gel solution with the concentration of 8% acrylamide and 0.06% bis-acrylamide (Bio-Rad, Hercules, CA, USA) were mixed and degas under vacuum for at least 20 min to remove oxygen. Then, 30 μl of 0.1 mg/mL ammonium persulfate (Sigma-Aldrich, St. Louis, MO, USA) and 20 μl TEMED (Applichem, Omaha, NE, USA) were added and placed into the mini protean casting strand and frame (Bio-Rad) to form 1-mm thickness of substrate. After letting the gel to polymerize for 30 to 45 min, gently remove and rinse gel with 50-mM HEPES, pH 8.5 (Applichem, Omaha, NE, USA). PA gel was then cut into circular shape with 14 mm diameters and placed in 24 well plates for the experiment. Sulfo-SANPAH (Pierce Biotechnologies, Rockford, IL USA) 0.5 mg/mL in 50-mM HEPES, pH 8.5 was pipetted onto the surface and exposed to the UV light for photoactivation procedure. After photoactivation, the substrate was washed several times in 50-mM HEPES. A 0.2 mg/mL of type I collagen (Sigma-Aldrich, St. Louis, MO, USA) was then layered onto the surface of gel and incubated 4 h at room temperature or overnight at 4°C on a shaker. After washing with PBS, the gels were stored in PBS at 4°C. Before platting the cells, the gel was exposed to UV for 15 min for the sterilization and replace PBS with complete culture medium for 1 h at 37°C.
Serial posts:
- Abstract : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- Background : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- Background : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- Methods : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- Methods : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- Methods : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [3]
- Methods : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [4]
- Results : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- Results : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- Discussion : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- Discussion : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- Conclusions : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- References : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- References : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- References : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [3]
- Acknowledgement : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- Author information : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [1]
- Author information : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions [2]
- Additional information : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- Additional file : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- Rights and permissions : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- About this article : Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions
- Figure 1. Cell attachment on tested materials. (A) Relative cell adherence of DFCs and dNC-PCs; (B) dental cells did little adhere on PA; representative pictures of DFCs. : Evaluation of implant
- Figure 2. Cell proliferation of dNC-PCs and DFCs on tested materials. (A) and (B) Relative cell numbers; (C) spheroid cell clusters on silicone (representative pictures for DFCs); Silicone (24 and 48 h). : Evaluation of implant
- Figure 3. Evaluation of programmed cell death (apoptosis) in dental stem cells. (A) Flow cytometry analyses (for details materials and methods) show percentage of vital cells (black number), apoptotic cells (blue number), and dead cells (red number). (B) Western blot analyses show the expression of the pro-apoptotic marker BAX and the anti-apoptotic marker BCL2. : Evaluation of implant
- Figure 4. Osteogenic differentiation of dental stem cells. Normalized ALP activity of dNC-PCs and DFCs on AP and SB (A) and on silicone (B). Cells were differentiated on standard cell culture dishes for control. : Evaluation of implant
- Figure 5. Evaluation of osteogenic differentiation. (A) Clustergram of PCR-array results; (B-C) histology of differentiated dental cells on AP (B) and SB (C). Representative results are shown for dNC-PCs. : Evaluation of implant
- Figure 6. Cultivation and osteogenic differentiation of DFCs on PA after modification with collagen I. (Left) Relative cell number and (Right) normalized ALP activity. : Evaluation of implant