Illustrating the potency of current Good Manufacturing Practice-compliant induced pluripotent stem cell lines as a source of multiple cell lineages using standardized protocols
| Autor: | Mahendra S. Rao, Thelma Y. Garcia, Ying Pei, Tomoko Hisai, Shereen Chew, Xianmin Zeng, Toshiharu Kasai, Hideki Taniguchi, Takanori Takebe, Deepak A. Lamba |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cancer Research Cell type Immunology Cell Induced Pluripotent Stem Cells Germ layer Tissue Banks Biology Retina Cell Line Mesoderm 03 medical and health sciences 0302 clinical medicine Neural Stem Cells medicine Immunology and Allergy Humans Cell Lineage Induced pluripotent stem cell Cell Engineering Genetics (clinical) Transplantation Retinal pigment epithelium Dopaminergic Neurons Mesenchymal stem cell Endothelial Cells Reproducibility of Results Cell Differentiation Cell Biology Reference Standards Neural stem cell Cell biology 030104 developmental biology medicine.anatomical_structure Oncology 030220 oncology & carcinogenesis Astrocytes Practice Guidelines as Topic Hepatocytes Guideline Adherence Cell bank |
| Zdroj: | Cytotherapy. 20(6) |
| ISSN: | 1477-2566 |
| Popis: | Background aims We have previously reported the generation of a current Good Manufacture Practice (cGMP)-compliant induced pluripotent stem cell (iPSC) line for clinical applications. Here we show that multiple cellular products currently being considered for therapy can be generated from a single master cell bank of this or any other clinically compliant iPSC line Methods Using a stock at passage 20 prepared from the cGMP-compliant working cell bank (WCB), we tested differentiation into therapeutically relevant cell types of the three germ layers using standardized but generic protocols. Cells that we generated include (i) neural stem cells, dopaminergic neurons and astrocytes; (ii) retinal cells (retinal pigment epithelium and photoreceptors); and (iii) hepatocyte, endothelial and mesenchymal cells. To confirm that these generic protocols can also be used for other iPSC lines, we tested the reproducibility of our methodology with a second clinically compliant line Results Our results confirmed that well-characterized iPSC lines have broad potency, and, despite allelic variability, the same protocols could be used with minimal modifications with multiple qualified lines. In addition, we introduced a constitutively expressed GFP cassette in Chr13 safe harbor site using a standardized previously described method and observed no significant difference in growth and differentiation between the engineered line and the control line indicating that engineered products can be made using a standardized methodology Conclusions We believe that our demonstration that multiple products can be made from the same WCB and that the same protocols can be used with multiple lines offers a path to a cost-effective strategy for developing cellular products from iPSC lines. |
| Databáze: | OpenAIRE |
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