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. 2021 May 6;16(6):1614–1628. doi: 10.1016/j.stemcr.2021.04.002

Figure 4.

Figure 4

Single hibernating HSCs can be manipulated by lentiviral transduction

(A) CD45+EPCR+CD48−CD150+ (ESLAM) cells were isolated and transduced with ZsGreen lentivirus and cultured together for 2 days in StemSpan supplemented with 10% fetal calf serum and IL-11. Cells were collected and virus was removed by collecting and re-sorting the cells into single wells and cultured in SCF-supplemented media for additional 10 days. A total of 4,001 total viable cells (a mixture of transduced and non-transduced cells) were re-sorted and transplanted into W41-CD45.1 (n = 6 recipients), and donor contribution and GFP expression were assessed by serial bleeds and flow cytometry analysis.

(B) Graph shows the percentage of clones surviving after 10 days post-addition of SCF, and the green bar indicated the percentage of GFP+ clones.

(C and D) Chimerism levels (20%–40%) were stable across all recipients at all time points (C), and an average of 1%–2% of donor cells was positive for GFP at 16 weeks post-transplantation (D). Bars show mean with SEM.