Generation of Induced Pluripotent Stem Cells from Human Testis Biopsy Derived Fibroblastic Cells

Although reported more than once, the generation of pluripotent stem cell populations from human testis remains questionable. We recently showed that the so - called human adult germline stem cells (haGSCs), which were claimed to be pluripotent, showed a gene expression profile comparable to human fibroblasts, suggesting that these cells are originated from a fibroblastic cells. We could derive human testicular fibroblast cells (hTFCs) from several routine testicular biopsies, naming them human testicular fibroblastic cells (hTFCs). The true conversion of unipotent mouse GSCs, also known as spermatogonial stem cells into pluripotent stem cells was recently shown. However, this conversion was not observed for human cells so far. We suggest that hTFCs isolated from routine biopsies can help in the understanding, modeling and also regenerative medicine for various diseases after reprogramming them to human induced pluripotent stem cells (iPS). We successfully reprogrammed hTFCs into human iPS cells by transducing them retrovirally with reprogramming factors as already demonstrated for human dermal fibroblasts. These hTFCs derived iPS cells are comparable with human embryonic stem cells from global gene expression, promoter methylation, and they show pluripotency judged by in vitro and in vivo differentiation. For transcriptome profiling, 400 ng of total DNA-free RNA was used as input for labelled cRNA synthesis (Illumina TotalPrep RNA Amplification Kit - Ambion) following the manufacturer's instructions (IVT: 12h). Quality-checked cRNA samples were hybridized as biological or technical duplicates for 18 h onto human-8 v3 expression BeadChips (Illumina), washed, stained, and scanned following guidelines and using materials/instrumentation supplied/suggested by the manufacturer. 8 samples were analyzed: NCL: NCL3 hESCs grown on feeders, bulk harvest at semi-sub confluence (1 replicate) HuES6: HuES6 hESCs grown on feeders, bulk harvest at semi-sub confluence (1 replicate) HT3: Human fibroblast-like testis cells, line 3 (1 replicate) HT16: Human fibroblast-like testis cells, line 16 (1 replicate) FS: FS human foreskin fibroblasts (1 replicate) BJ: BJ human fibroblasts (1 replicate) iPS3: hiPSC line 2 derived from HT3 (1 replicate) iPS16: hiPSC line 2 derived from HT16 (1 replicate)