The chondrogenic differentiation potential of PDLSCs was detected using Alcian Blue staining. In brief, PDLSCs were collected in 15-mL centrifuge tubes (Invitrogen) at a density of 2 × 105 cells/tube and then cultured with chondrogenic induction medium (Cyagen) until those cells aggregated and formed a suspending cell cluster. The medium was refreshed every 2 days. After a 21-day chondrogenic induction, the cell clusters were fixed with 4% paraformaldehyde, and the chondrogenic differentiation potential of PDLSCs was evaluated using Alcian Blue solution (Cyagen) according to previous studies (13 (link), 37 (link)).
15 ml centrifuge tubes
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The 15-mL centrifuge tubes are a common laboratory equipment used for various sample preparation and analysis procedures. These tubes are designed to hold up to 15 milliliters of liquid or solid samples and are suitable for a wide range of centrifugation applications.
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4 protocols using 15 ml centrifuge tubes
1
Chondrogenic Differentiation of PDLSCs
2
Chondrogenic Induction of PDLSCs
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For chondrogenic induction, P4 cells were collected in 15-mL centrifuge tubes (Invitrogen) at a density of 2 × 105 cells/tube. The cells were washed with PBS and then resuspended in chondro-inductive medium (Cyagen). After 24 h, cell spheres were visible at the bottom of the tubes. The medium was exchanged every 2 days.
According to studies reported [30 (link)], Alcian blue staining was performed after chondrogenic induction for 21 days to measure the chondrogenic differentiation ability of PDLSCs.
According to studies reported [30 (link)], Alcian blue staining was performed after chondrogenic induction for 21 days to measure the chondrogenic differentiation ability of PDLSCs.
3
Plant Tissue RNA Extraction Protocol
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Excess moisture was removed from the collected fresh tissue by blotting the tissue with dry paper towels, then the tissue was transferred to 15 mL centrifuge tubes (Thermo Fisher Scientific), flash frozen with liquid N2, and stored at -80°C. About 1 g of harvested plant tissue was pulverized under liquid N2 in an RNase-free mortar and pestle. RNA was extracted with TRIzol Reagents (Thermo Fisher Scientific), followed by treatment with DNase I (QIAGEN, Hilden, Germany) and clean-up with RNeasy Mini Kit (QIAGEN). The quality of RNA was analyzed with an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). The RNA integrity number (RIN) of all samples was ≥7.8.
4
Cigarette Smoke Extract Preparation
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Approximately 30–50 mL of cigarette smoke was drawn into the syringe and bubbled into sterile PBS in 15 mL centrifuge tubes (Thermo Fisher) to prepare cigarette smoke (CS) extract. We used one cigarette for the preparation of 10 mL of solution. To remove insoluble particles, the CS extract solution was filtered (0.22 μm; Merck Millipore) and stored for further use.
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