Using low-risk factors to generate non-integrated human induced pluripotent stem cells from urine-derived cells

Table 1 Induction efficiency and characterization of iPSCs induced from hUCs using the 6F/BM1-4C system

Donor	Age	Gender	Cell number	Death rate^a (%)	Efficiency (%)	No.	Non-integration	Karyotype	FACS	BSP	Teratoma^b	EB formation	Immunofluorescent
1	23	Female	3 × 10⁶	53.8	0.0468	UC1iPSC1	+^c	+	+	+	+	+	+
1	23	Female	3 × 10⁶	53.8	0.0468	UC1iPSC2	+^c	+	+	–	–	–	–
2	26	Male	2.8 × 10⁶	80.37	0.0015	UC2iPSC1	+^c	+	+	–	+	+	+
2	26	Male	2.8 × 10⁶	80.37	0.0015	UC2iPSC2	+^c	+		+	–	–	–
3	27	Male	3 × 10⁶	50	0.0741	UC3iPSC1	+^c	+	–	–	+	+	–
3	27	Male	3 × 10⁶	50	0.0741	UC3iPSC2	+^c	+	–	+	–	–	–
4	26	Female	3 × 10⁶	71	0.00089	UC4iPSC1	+^c	+	–	–	–	–	–
4	26	Female	3 × 10⁶	71	0.00089	UC4iPSC2	+^c	+	–	+^e	–	–	–
5	24	Male	3 × 10⁶	66.35	0.00095	UC5iPSC1	+^c	+	–	–	+	–	–
5	24	Male	3 × 10⁶	66.35	0.00095	UC5iPSC2	+^c	+	–	+^e	–	–	–
6	30	Female	2.53 × 10⁶	36	0.00021	UC6iPSC1	+^d	+	–	–	–	–	–
6	30	Female	2.53 × 10⁶	36	0.00021	UC6iPSC2	+^c	+	–	+	–	–	–
7	29	Male	3 × 10⁶	27.5	0.0429	UC7iPSC1	+^c	+	–	+^e	–	–	–
7	29	Male	3 × 10⁶	27.5	0.0429	UC7iPSC2	+^c	+	–	–	–	–	–

At nucleofection, the hUC passage number was 2
+ iPSCs identified, – characterization not identified, iPSC induced pluripotent stem cell, hUC human urine-derived cell, FACS fluorescence-activated cell sorting, EB embryoid body, HE hematoxylin and eosin
^aTaking supernatant (containing nonadherent cells) to count cells on the 3rd day after electronic transformation, the amount of counted cells was the amount of dead cells. Death rate calculated as: death rate = amount of dead cells/amount of cells for electroporation
^bWe injected five different donor-derived iPSCs in the teratoma experiment. Each NOD/SCID mouse was injected with a cell. In 6–8 weeks, three NOD/SCID mice formed teratomas and were dissected and HE stained. In the 8th week, one NOD/SCID mice had granule-shaped teratoma, and the teratoma was taken out and HE stained in the 12th week. One mouse did not form teratoma by observation
^cExogenous gene sequence did not integrate in the genome
^dExogenous gene sequence integrated in the genome
^eCorresponding primary cells did not detect the pluripotency gene promoter DNA methylation due to the exhausted cells

ISSN: 1757-6512