Oct 11, 2015
Medium change for qHAES
Period: 5, 4, 3, 2, 2
Maintenance
Hep(C)iPS#25
Hep2055-iPS#10
pHAES06: two dishes.
Accutase treatment to iPSCs (iPSellon) versus liberase for HAES.
1 x 10^4 cells per well in a 96-well dish are cultivated for EB formation.
EB formation in the EB medium for 10 days.
EB medium (the EB (iPSellon w/o FGF2, 10 days, Nunclon))
Knockout DMEM (380 ml)
KSR (100 ml)
Pc/SM (5 ml)
GlutaMax (5 ml)
Sodium Pyruvate (5 ml)
MEM (5 ml)
beta-ME (0.5 ml)
Plate 20 EBs in a 24-well dish in XF32 with H, I, bF for 2 to 3 weeks.
Two lines from one patient (Hep2055-iPS#10 and #32)
---Elimination of Sendai virus was successful.
Hep(C)iPS: Fourteen lines from another patient
(iPS#24, Hep(C)-iPS#12, #19, and #25 are on; #62, #66, #100, #53, #41, #116: off)
---Sendai virus of Hep(C)-iPS#12, #19, and #25 were successfully eliminated.
---iPS#24 cells were differentiated into hepatocytes by the Saeko method.
Exome analysis and karyotypic analysis will be performed.
in vivo (kidney assay) and in vitro assay (paraffin-embedded slides).
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