Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system
Introduction: Genetically modified human induced pluripotent stem cell (iPSC)-based regenerative medicine has substantial potential in the treatment of refractory human diseases. Thus, preclinical studies on the safety and efficacy of these products are essential. Non-human primate (NHP) models such...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-12-01
|
| Series: | Regenerative Therapy |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320424000269 |
| _version_ | 1797256321886584832 |
|---|---|
| author | Masahiro Tanaka Yoshihiro Iwamoto Bo Wang Eri Imai Munehiro Yoshida Shoichi Iriguchi Shin Kaneko |
| author_facet | Masahiro Tanaka Yoshihiro Iwamoto Bo Wang Eri Imai Munehiro Yoshida Shoichi Iriguchi Shin Kaneko |
| author_sort | Masahiro Tanaka |
| collection | DOAJ |
| description | Introduction: Genetically modified human induced pluripotent stem cell (iPSC)-based regenerative medicine has substantial potential in the treatment of refractory human diseases. Thus, preclinical studies on the safety and efficacy of these products are essential. Non-human primate (NHP) models such as the rhesus macaque are highly similar to humans in terms of size, lifespan, and immune system, rendering them superior models. However, effective gene transduction in rhesus macaque iPSCs (Rh-iPSCs) remains challenging. In this study, we investigated the effective gene transduction into Rh-iPSCs and its effect on differentiation efficiency. Methods: We established a gene transduction method using the piggyBac transposon vector system. Gene transduced Rh-iPSCs were analyzed for undifferentiated markers. We did teratoma assay to check pluripotency. Gene transduced Rh-iPSCs were differentiated into hematopoietic stem and progenitor cells (HSPCs) and T-cell lineage cells. Additionally, gene transduced Rh-iPSCs were compared the differentiation efficiency with parental Rh-iPSCs. Results: We could establish a gene transduction method using the piggyBac transposon vector system, demonstrating high efficiency and stable transgene expression in Rh-iPSCs. These Rh-iPSCs maintained long-term gene expression while expressing undifferentiated markers. Teratoma assay indicated that these Rh-iPSCs had pluripotency. These Rh-iPSCs could differentiate into HPSCs and T cells that express transgenes. These Rh-iPSCs can differentiate into hematopoietic stem cells and T cells that express transgenes. No significant differences in efficiency of differentiation were observed between parental Rh-iPSCs and these Rh-iPSCs. Conclusions: These results indicate that the piggyBac transposon vector is an excellent gene transfer tool for rhesus macaque iPSCs and could contribute to the advancement of preclinical studies using rhesus macaque iPSCs. |
| first_indexed | 2024-04-24T22:19:54Z |
| format | Article |
| id | doaj.art-1ddaf5267efa4756bc597686a33ece9a |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-04-24T22:19:54Z |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-1ddaf5267efa4756bc597686a33ece9a2024-03-20T06:09:31ZengElsevierRegenerative Therapy2352-32042024-12-0127104111Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac systemMasahiro Tanaka0Yoshihiro Iwamoto1Bo Wang2Eri Imai3Munehiro Yoshida4Shoichi Iriguchi5Shin Kaneko6Shin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, JapanPrincess Margaret Cancer Centre, University Health Network, Toronto, Ontario, CanadaShin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, JapanShin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, JapanShin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, JapanShin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, JapanShin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan; Corresponding author. Shin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.Introduction: Genetically modified human induced pluripotent stem cell (iPSC)-based regenerative medicine has substantial potential in the treatment of refractory human diseases. Thus, preclinical studies on the safety and efficacy of these products are essential. Non-human primate (NHP) models such as the rhesus macaque are highly similar to humans in terms of size, lifespan, and immune system, rendering them superior models. However, effective gene transduction in rhesus macaque iPSCs (Rh-iPSCs) remains challenging. In this study, we investigated the effective gene transduction into Rh-iPSCs and its effect on differentiation efficiency. Methods: We established a gene transduction method using the piggyBac transposon vector system. Gene transduced Rh-iPSCs were analyzed for undifferentiated markers. We did teratoma assay to check pluripotency. Gene transduced Rh-iPSCs were differentiated into hematopoietic stem and progenitor cells (HSPCs) and T-cell lineage cells. Additionally, gene transduced Rh-iPSCs were compared the differentiation efficiency with parental Rh-iPSCs. Results: We could establish a gene transduction method using the piggyBac transposon vector system, demonstrating high efficiency and stable transgene expression in Rh-iPSCs. These Rh-iPSCs maintained long-term gene expression while expressing undifferentiated markers. Teratoma assay indicated that these Rh-iPSCs had pluripotency. These Rh-iPSCs could differentiate into HPSCs and T cells that express transgenes. These Rh-iPSCs can differentiate into hematopoietic stem cells and T cells that express transgenes. No significant differences in efficiency of differentiation were observed between parental Rh-iPSCs and these Rh-iPSCs. Conclusions: These results indicate that the piggyBac transposon vector is an excellent gene transfer tool for rhesus macaque iPSCs and could contribute to the advancement of preclinical studies using rhesus macaque iPSCs.http://www.sciencedirect.com/science/article/pii/S2352320424000269iPSCRhesus macaquepiggyBacT-cell differentiation |
| spellingShingle | Masahiro Tanaka Yoshihiro Iwamoto Bo Wang Eri Imai Munehiro Yoshida Shoichi Iriguchi Shin Kaneko Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system Regenerative Therapy iPSC Rhesus macaque piggyBac T-cell differentiation |
| title | Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system |
| title_full | Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system |
| title_fullStr | Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system |
| title_full_unstemmed | Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system |
| title_short | Effective and stable gene transduction in rhesus macaque iPSCs capable of T-lineage differentiation utilizing the piggyBac system |
| title_sort | effective and stable gene transduction in rhesus macaque ipscs capable of t lineage differentiation utilizing the piggybac system |
| topic | iPSC Rhesus macaque piggyBac T-cell differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S2352320424000269 |
| work_keys_str_mv | AT masahirotanaka effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT yoshihiroiwamoto effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT bowang effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT eriimai effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT munehiroyoshida effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT shoichiiriguchi effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem AT shinkaneko effectiveandstablegenetransductioninrhesusmacaqueipscscapableoftlineagedifferentiationutilizingthepiggybacsystem |