A feeder- and xeno-free human induced pluripotent stem cell line obtained from primary human dermal fibroblasts with epigenetic repression of reprogramming factors expression: GPCCi001-A

• Published in: Stem cell research Vol. 20; no. C; pp. 34 - 37
• Main Authors: Lach, Michał Stefan, Wroblewska, Joanna P., Augustyniak, Ewelina, Kulcenty, Katarzyna, Suchorska, Wiktoria Maria
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.04.2017; Elsevier
• Subjects: Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Cellular Reprogramming; Embryoid Bodies - metabolism; Embryoid Bodies - pathology; Epigenetic Repression; Feeder Cells - cytology; Female; Fibroblasts - cytology; Fibroblasts - metabolism; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Karyotype; Microscopy, Fluorescence; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2017.02.004

The primary human dermal fibroblasts (PHDFs) from breast cancer patient were obtained to generate the human induced pluripotent stem cell line GPCCi001-A via lentiviral transfection. Thus, a modified EF1a-hSTEMCCA-loxP with tetO operator which regulates transgene expression was used. This method takes advantage of epigenetic regulation of transcription and allows for stable silencing of the reprogramming factors in obtained hiPS cells. To increase the potential utility of hiPSCs for clinical applications, they were adapted to feeder- and xeno-free conditions. The pluripotency of GPCCi001-A cell line and ability to differentiate into three germ layers was confirmed.