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. 2020 Dec 19;23:629–639. doi: 10.1016/j.omtn.2020.12.012

Figure 3.

Figure 3

Transfer of a HAC into human iPSCs for biomedical application

(A) Outline of the chromosome transfer of the modified 21HAC2 from CHO cells to T-iPSCs and manipulations of the basal-HAC in T-iPSCs. (B) Fluorescence microscopy images of basal-HAC (left) and KO1-HAC (right) T-iPSCs on MEFs. Phase, phase contrast; GFP, enhanced green fluorescent protein; KO, Kusabira Orange fluorescent protein. (C) FISH analysis of T-iPSCs containing KO1-HAC. Blue, DAPI; red, alpha satellite probe for chromosomes 13 and 21 and HAC (p11-4); green, plasmid containing KO1 (ΦC31 attB-NeoR-pUbc-KO1). (D) Flow cytometry data of KO1-HAC T-iPSCs. Expression of fluorescent proteins in all live cells and pluripotency markers SSEA4 and TRA1-60 in GFP+KO+ cells are shown. (E) Images of an iPSC-Sac containing hematopoietic progenitor cells on 10T1/2 feeder cells. (F) Flow cytometry data of trypsinized sac cells derived from KO1-HAC T-iPSCs. Expression of fluorescent proteins in all live cells (left) and hematopoietic markers CD34 and CD43 in GFP+KO+ cells (right) are shown. (G) Expression of CD3 on T cells differentiated from KO-inserted basal-HAC T-iPSCs.