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Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications

Title: Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications
Authors: Sergey A. Sinenko; Elena V. Skvortsova; Mikhail A. Liskovykh; Sergey V. Ponomartsev; Andrey A. Kuzmin; Aleksandr A. Khudiakov; Anna B. Malashicheva; Natalia Alenina; Vladimir Larionov; Natalay Kouprina; Alexey N. Tomilin
Source: Cells, Vol 7, Iss 12, p 261 (2018)
Publisher Information: MDPI AG
Publication Year: 2018
Collection: Directory of Open Access Journals: DOAJ Articles
Subject Terms: human artificial chromosome (HAC); alphoid tetO -HAC; induced pluripotent stem cells (iPSCs); microcell-mediated chromosome transfer (MMCT); cell reprogramming; Cytology; QH573-671
Description: Alphoid tetO -type human artificial chromosome (HAC) has been recently synthetized as a novel class of gene delivery vectors for induced pluripotent stem cell (iPSC)-based tissue replacement therapeutic approach. This HAC vector was designed to deliver copies of genes into patients with genetic diseases caused by the loss of a particular gene function. The alphoid tetO -HAC vector has been successfully transferred into murine embryonic stem cells (ESCs) and maintained stably as an independent chromosome during the proliferation and differentiation of these cells. Human ESCs and iPSCs have significant differences in culturing conditions and pluripotency state in comparison with the murine naïve-type ESCs and iPSCs. To date, transferring alphoid tetO -HAC vector into human iPSCs (hiPSCs) remains a challenging task. In this study, we performed the microcell-mediated chromosome transfer (MMCT) of alphoid tetO -HAC expressing the green fluorescent protein into newly generated hiPSCs. We used a recently modified MMCT method that employs an envelope protein of amphotropic murine leukemia virus as a targeting cell fusion agent. Our data provide evidence that a totally artificial vector, alphoid tetO -HAC, can be transferred and maintained in human iPSCs as an independent autonomous chromosome without affecting pluripotent properties of the cells. These data also open new perspectives for implementing alphoid tetO -HAC as a gene therapy tool in future biomedical applications.
Document Type: article in journal/newspaper
Language: English
Relation: https://www.mdpi.com/2073-4409/7/12/261; https://doaj.org/toc/2073-4409; https://doaj.org/article/fe320cdbeceb4510ae59b59ce446545f
DOI: 10.3390/cells7120261
Availability: https://doi.org/10.3390/cells7120261; https://doaj.org/article/fe320cdbeceb4510ae59b59ce446545f
Accession Number: edsbas.FAC8A34E
Database: BASE