Methods of reprogramming animal somatic cells

A method comprising

• (a) contacting an isolated mammalian somatic cell or a nucleus thereof with a cell-free extract
• (i) obtained from undifferentiated cells engineered to overexpress one or more reprogramming factors and/or
• (ii) supplemented with one or more reprogramming factors,
• wherein the reprogramming factors are selected from the group consisting of hsa-miR-18a, hsa-miR-18b, hsa-miR-20b, hsa-miR-96, hsa-miR-106a, hsa-miR-107, hsa-miR-141, hsa-miR-183, hsa-miR-187, hsa-miR-203, hsa-miR-211, hsa-miR-217, hsa-miR-218-1, hsa-miR-218-2, hsa-miR-302a, hsa-miR-302c, hsa-miR-302d, hsa-miR-330, hsa-miR-363, hsa-miR-367, hsa-miR-371, hsa-miR-372, hsa-miR-373, hsa-miR-496, hsa-miR-50B, hsa-miR-512-3p, hsa-miR-512-5p, hsa-miR-515-3p, hsa-miR-515-5p, hsa-miR-516-5p, hsa-miR-517, hsa-miR-517a, hsa-miR-518b, hsa-miR-51Bc, hsa-miR-518e, hsa-miR-51ge, hsa-miR-520a, hsa-miR-520b, hsa-miR-520e, hsa-miR-520g, hsa-miR-520h, hsa-miR-523, hsa-miR-524, hsa-miR-525, hsa-miR-526a-1, and hsa-miR-526a-2;
• and
• (b) analyzing the karyotype and/or the extent of reprogramming of the mammalian cell or nucleus thereof produced by step(a) by
• (i) karyotyping the mammalian cell or nucleus thereof by chromosome spread, spectral karyotyping, or total genomic hybridization assay, or
• (ii) determining whether the cell or nucleus thereof expresses
• (1) E-cadherin mRNA at levels of at least 5% of the expression level of the housekeeping gene GAPD;
• and/or
• (2) LIN28expression at levels of at least 5% of the housekeeping gene GAPD.

A method comprising

• (a) contacting an isolated mammalian somatic cell or a nucleus thereof with a cell-free extract,
• (i) from human embryonic stem cells, engineered to overexpress one or more reprogramming factors and/or
• (ii) supplemented with one or more reprogramming factors,
• wherein the reprogramming factors are selected from the group consisting of BARX1, CROC4, DNMT3B, H2AFX, HHEX, HISTIH2AB, HISTIH4J, HMGB2, hsa-miR-18a, hsa-miR-18b, hsa-miR-20b, hsa-miR-96, hsa-miR-106a, hsa-miR-107, hsa-miR-141, hsa-miR-183, hsa-miR-187, hsa-miR-203, hsa-miR-211, hsa-miR-217, hsa-miR-218-1, hsa-miR-218-2, hsa-miR-302a, hsa-miR-302c, hsa-miR-302d, hsa-miR-330, hsa-miR-363, hsa-miR-367, hsa-miR-371, hsa-miR-372, hsa-miR-373, hsa-miR-496, hsa-miR-50B, hsa-miR-512-3p, hsa-miR-512-5p, hsa-miR-515-3p, hsa-miR-515-5p, hsa-miR-516-5p, hsa-miR-517, hsa-miR-517a, hsa-miR-518b, hsa-miR-51Bc, hsa-miR-518e, hsa-miR-51ge, hsa-miR-520a, hsa-miR-520b, hsa-miR-520e, hsa-miR-520g, hsa-miR-520h, hsa-miR-523, hsa-miR-524, hsa-miR-525, hsa-miR-526a-1, hsa-miR-526a-2, LEFTB, LHX1, LHX6, LIN28, MYBL2, MYC, MYCN, NANOG, NFIX, OCT3/4(POU5F1), OCT6 (POU3F1), OTX2, PHC1, SALL4, SOX2, TERF1, TERT, TGIF, VENTX2, ZIC2, ZIC3, ZIC5, and ZNF206;
• and
• (b) analyzing the karyotype and/or the extent of reprogramming of the mammalian cell or nucleus thereof produced by step (a) by
• (i) karyotyping the mammalian cell or nucleus thereof by chromosome spread, spectral karyotyping, or total genomic hybridization assay, or
• (ii) determining whether the cell or nucleus thereof expresses
• (1) E-cadherin mRNA at levels of at least 5% of the expression level of the housekeeping gene GAPD;
• and/or
• (2) LIN28 expression at levels of at least 5% of the housekeeping gene GAPD.