A Simplified Method for the Aspiration of Bone Marrow from Patients Undergoing Hip and Knee Joint Replacement for Isolating Mesenchymal Stem Cells and In Vitro Chondrogenesis

Table 1

Reconstitution of incomplete chondrogenesis medium.

Reagent	Source	Stock solution (storage)	Final conc.	Mix and filter sterilize
DMEM with high glucose, 1x	D5671; (Sigma-Aldrich)		1x	475 mL
L-Ascorbic acid 2-phosphate trisodium salt	323-44822 (Wako)	5 mg/mL in dH2O (4°C)	50 μg/mL	5 mL
L-Proline	P5607; (Sigma-Aldrich)	50 mg/mL in dH2O (4°C)	50 μg/mL	500 μL
Insulin-transferrin-selenium-sodium pyruvate (ITS-A) (100x)	51300-044; (Gibco by Life Technologies)		1x	5 mL
Sodium pyruvate (100 mM)	11360-070; (Gibco by Life Technologies)		1 mM	5 mL
Penicillin-streptomycin; (10,000 U/mL)	15140-122; (Gibco by Life Technologies)		100 U/mL	5 mL
GlutaMAX supplement (100x)	35050-061; (Gibco by Life Technologies)		1x	5 mL
Linoleic acid; (1 gm/mL)	L1012; (Sigma-Aldrich)	100 μg/μL  dilute 5 μL (1 gm/mL) to 50 μL in dH2O	5.33 μg/mL	26.7 μL
Dexamethasone; (MW: 392.46)	D4902, (Sigma-Aldrich)	Solution-A (10 mM) (dissolve 25 mg in 6.37 mL EtOH and store at −20°C); Solution-B (1 mM): dilute 100 μL solution-A to 1 mL in dH2O	100 nM	50 μL (solution-B)