Efficient and nontoxic biomolecule delivery to primary human hematopoietic stem cells using nanostraws
(2020) In Proceedings of the National Academy of Sciences of the United States of America 117(35). p.21267-21273- Abstract
Introduction of exogenous genetic material into primary stem cells is essential for studying biological function and for clinical applications. Traditional delivery methods for nucleic acids, such as electroporation, have advanced the field, but have negative effects on stem cell function and viability. We introduce nanostraw-assisted transfection as an alternative method for RNA delivery to human hematopoietic stem and progenitor cells (HSPCs). Nanostraws are hollow alumina nanotubes that can be used to deliver biomolecules to living cells. We use nanostraws to target human primary HSPCs and show efficient delivery of mRNA, short interfering RNAs (siRNAs), DNA oligonucleotides, and dextrans of sizes ranging from 6 kDa to 2,000 kDa.... (More)
Introduction of exogenous genetic material into primary stem cells is essential for studying biological function and for clinical applications. Traditional delivery methods for nucleic acids, such as electroporation, have advanced the field, but have negative effects on stem cell function and viability. We introduce nanostraw-assisted transfection as an alternative method for RNA delivery to human hematopoietic stem and progenitor cells (HSPCs). Nanostraws are hollow alumina nanotubes that can be used to deliver biomolecules to living cells. We use nanostraws to target human primary HSPCs and show efficient delivery of mRNA, short interfering RNAs (siRNAs), DNA oligonucleotides, and dextrans of sizes ranging from 6 kDa to 2,000 kDa. Nanostraw-treated cells were fully functional and viable, with no impairment in their proliferative or colony-forming capacity, and showed similar long-term engraftment potential in vivo as untreated cells. Additionally, we found that gene expression of the cells was not perturbed by nanostraw treatment, while conventional electroporation changed the expression of more than 2,000 genes. Our results show that nanostraw-mediated transfection is a gentle alternative to established gene delivery methods, and uniquely suited for nonperturbative treatment of sensitive primary stem cells.
(Less)
- author
- Schmiderer, Ludwig LU ; Subramaniam, Agatheeswaran LU ; Žemaitis, Kristijonas LU ; Bäckström, Alexandra LU ; Yudovich, David LU ; Soboleva, Svetlana LU ; Galeev, Roman LU ; Prinz, Christelle N. LU ; Larsson, Jonas LU and Hjort, Martin LU
- organization
-
- Division of Molecular Medicine and Gene Therapy
- StemTherapy: National Initiative on Stem Cells for Regenerative Therapy
- Stem Cell Metabolism (research group)
- LUCC: Lund University Cancer Centre
- Solid State Physics
- NanoLund: Centre for Nanoscience
- WCMM-Wallenberg Centre for Molecular Medicine
- Chemical Biology and Therapeutics (research group)
- publishing date
- 2020
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Electroporation, Hematopoietic stem cell, MRNA delivery, Nanostraws, Transfection
- in
- Proceedings of the National Academy of Sciences of the United States of America
- volume
- 117
- issue
- 35
- pages
- 7 pages
- publisher
- National Academy of Sciences
- external identifiers
-
- scopus:85090508841
- pmid:32817519
- ISSN
- 0027-8424
- DOI
- 10.1073/pnas.2001367117
- language
- English
- LU publication?
- yes
- id
- 379b6603-366e-4f5b-8d90-ba1ab091cb8d
- date added to LUP
- 2020-09-30 10:40:04
- date last changed
- 2024-08-08 01:53:05
@article{379b6603-366e-4f5b-8d90-ba1ab091cb8d, abstract = {{<p>Introduction of exogenous genetic material into primary stem cells is essential for studying biological function and for clinical applications. Traditional delivery methods for nucleic acids, such as electroporation, have advanced the field, but have negative effects on stem cell function and viability. We introduce nanostraw-assisted transfection as an alternative method for RNA delivery to human hematopoietic stem and progenitor cells (HSPCs). Nanostraws are hollow alumina nanotubes that can be used to deliver biomolecules to living cells. We use nanostraws to target human primary HSPCs and show efficient delivery of mRNA, short interfering RNAs (siRNAs), DNA oligonucleotides, and dextrans of sizes ranging from 6 kDa to 2,000 kDa. Nanostraw-treated cells were fully functional and viable, with no impairment in their proliferative or colony-forming capacity, and showed similar long-term engraftment potential in vivo as untreated cells. Additionally, we found that gene expression of the cells was not perturbed by nanostraw treatment, while conventional electroporation changed the expression of more than 2,000 genes. Our results show that nanostraw-mediated transfection is a gentle alternative to established gene delivery methods, and uniquely suited for nonperturbative treatment of sensitive primary stem cells.</p>}}, author = {{Schmiderer, Ludwig and Subramaniam, Agatheeswaran and Žemaitis, Kristijonas and Bäckström, Alexandra and Yudovich, David and Soboleva, Svetlana and Galeev, Roman and Prinz, Christelle N. and Larsson, Jonas and Hjort, Martin}}, issn = {{0027-8424}}, keywords = {{Electroporation; Hematopoietic stem cell; MRNA delivery; Nanostraws; Transfection}}, language = {{eng}}, number = {{35}}, pages = {{21267--21273}}, publisher = {{National Academy of Sciences}}, series = {{Proceedings of the National Academy of Sciences of the United States of America}}, title = {{Efficient and nontoxic biomolecule delivery to primary human hematopoietic stem cells using nanostraws}}, url = {{http://dx.doi.org/10.1073/pnas.2001367117}}, doi = {{10.1073/pnas.2001367117}}, volume = {{117}}, year = {{2020}}, }