Isolation and Culture of Human Bone Marrow Stem Cells

This study was approved by the Institutional Review Board of Yonsei University College of Medicine (IRB no. 4-2017-0232). Human bone marrow aspirates were obtained from the point 3 cm posterior to the anterior superior iliac spine of 10 healthy adult donors with a mean age of 35 years (range: 30–40 years). hBMSCs were maintained in low-glucose Dulbecco’s modified Eagle’s medium (DMEM-LG; Gibco BRL, Rockville, MD, USA) with 10% fetal bovine serum (FBS; Gibco) and 1% antibiotic-antimycotic solution (Gibco). Human embryonic kidney 293 (HEK293) and 293FT cells were obtained from American Type Culture Collection (Manassas, VA, USA). These cells were maintained in high-glucose DMEM (DMEM-HG; Gibco) containing 10% FBS and 1% antibiotic-antimycotic solution (Gibco). DKK1 (R&D Systems) was used at a concentration of 0.1 mg/mL in hBMSCs^{53 (link)}. Wnt3a (cat. no. 5036-WN; R&D Systems) was used at a concentration of 10 ng/mL in hBMSCs^{54 (link),55 (link)}.