Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications
Alphoid<sup>tetO</sup>-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 patie...
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doaj-fe320cdbeceb4510ae59b59ce446545f2020-11-24T20:44:36ZengMDPI AGCells2073-44092018-12-0171226110.3390/cells7120261cells7120261Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical ApplicationsSergey A. Sinenko0Elena V. Skvortsova1Mikhail A. Liskovykh2Sergey V. Ponomartsev3Andrey A. Kuzmin4Aleksandr A. Khudiakov5Anna B. Malashicheva6Natalia Alenina7Vladimir Larionov8Natalay Kouprina9Alexey N. Tomilin10Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg 194064, RussiaInstitute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg 194064, RussiaDevelopmental Therapeutics Branch, National Cancer Institute, Bethesda, MD 20892, USAInstitute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg 194064, RussiaInstitute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg 194064, RussiaAlmazov National Medical Research Centre, 2 Akkuratova Str., St-Petersburg 197341, RussiaAlmazov National Medical Research Centre, 2 Akkuratova Str., St-Petersburg 197341, RussiaMax-Delbruck Center for Molecular Medicine, 10 Robert-Rössle-Straße, 13125 Berlin, GermanyDevelopmental Therapeutics Branch, National Cancer Institute, Bethesda, MD 20892, USADevelopmental Therapeutics Branch, National Cancer Institute, Bethesda, MD 20892, USAInstitute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg 194064, RussiaAlphoid<sup>tetO</sup>-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<sup>tetO</sup>-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<sup>tetO</sup>-HAC vector into human iPSCs (hiPSCs) remains a challenging task. In this study, we performed the microcell-mediated chromosome transfer (MMCT) of alphoid<sup>tetO</sup>-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<sup>tetO</sup>-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<sup>tetO</sup>-HAC as a gene therapy tool in future biomedical applications.https://www.mdpi.com/2073-4409/7/12/261human artificial chromosome (HAC)alphoid<sup>tetO</sup>-HACinduced pluripotent stem cells (iPSCs)microcell-mediated chromosome transfer (MMCT)cell reprogramming |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
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 |
spellingShingle |
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 Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications Cells human artificial chromosome (HAC) alphoid<sup>tetO</sup>-HAC induced pluripotent stem cells (iPSCs) microcell-mediated chromosome transfer (MMCT) cell reprogramming |
author_facet |
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 |
author_sort |
Sergey A. Sinenko |
title |
Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications |
title_short |
Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications |
title_full |
Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications |
title_fullStr |
Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications |
title_full_unstemmed |
Transfer of Synthetic Human Chromosome into Human Induced Pluripotent Stem Cells for Biomedical Applications |
title_sort |
transfer of synthetic human chromosome into human induced pluripotent stem cells for biomedical applications |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2018-12-01 |
description |
Alphoid<sup>tetO</sup>-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<sup>tetO</sup>-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<sup>tetO</sup>-HAC vector into human iPSCs (hiPSCs) remains a challenging task. In this study, we performed the microcell-mediated chromosome transfer (MMCT) of alphoid<sup>tetO</sup>-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<sup>tetO</sup>-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<sup>tetO</sup>-HAC as a gene therapy tool in future biomedical applications. |
topic |
human artificial chromosome (HAC) alphoid<sup>tetO</sup>-HAC induced pluripotent stem cells (iPSCs) microcell-mediated chromosome transfer (MMCT) cell reprogramming |
url |
https://www.mdpi.com/2073-4409/7/12/261 |
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