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Efficient and nontoxic biomolecule delivery to primary human hematopoietic stem cells using nanostraws

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 orcid (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.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
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
2024-04-17 15:46:43
@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}},
}