Extended Data Fig. 4: In vitro functional assay of YSMPs.
From: Deciphering human macrophage development at single-cell resolution
a, Gating strategy for sorting of the YSMPs (CD45+CD34+CD44+) from a CS12 yolk sac. b, Representative morphologies of bulk cultures (100 cells per well) of negative control cells (CD45+CD34−CD44−, n = 5 replication wells) and YSMPs (CD45+CD34+CD44+, n = 16 replication wells) after 14 days of culture on MS5 feeder layer. n = 3 independent experiments from one sample of CS11 yolk sac and two samples of CS 12 yolk sac for YSMPs. Scale bars, 100 μm. c, Representative FACS analysis of cells collected from bulk cultures of YSMPs. Note that the myeloid cells (CD33+) are predominant, in contrast to a small number of erythroid cells (CD235a+) detected (n = 3 independent experiments). d, Representative morphologies of haematopoietic cells generated by a single YSMP from a CS13 yolk sac after 3 and 10 days of culture on MS5 feeder layer. In total, 184 YSMPs were individually cultured and 67 of them generated morphologically typical haematopoietic clusters. Scale bars, 100 μm. e, Representative FACS analysis of four kinds of distinct differentiation potential of single YSMPs. Cells were collected from the single-cell YSMP cultures and in total 39 wells were individually analysed. Lineage differentiation potentials are indicated in red for each clone. Mo/Mac, monocytes/macrophages (CD45+CD33+CD14+); Gr, granulocytes (CD45+CD33+CD66b+); Ery, erythrocytes (CD235a+); Mk, megakaryocytes (CD41a+).
