Isolation and Analysis of CD34+ Cells

BM samples were collected from two adult healthy donors and two AA patients at the First Affiliated Hospital of Zhejiang Chinese University. All participants provided written informed consent before enrollment in this study. Biospecimen collection protocols complied with local guidelines and were approved by the Ethics Committee of the First Affiliated Hospital of Zhejiang Chinese University. The isolation procedure was described previously (Macosko et al., 2015 (link)). Mononuclear cells (MNCs) were isolated using Ficoll–Hypaque gradient separation (Tianjin Hao Yang Biological Products Technology, China). CD34 ⁺ cells were purified from MNCs with the human anti-CD34 MicroBeads Isolation Kit (Miltenyi Biotec) according to the manufacturer’s specifications, and CD34 ⁺ cells were obtained. Then, MNCs mixed with CD34 ⁺ cells at a 4:1 ratio were analyzed by 10× Genomics.