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Mouse and human HSPC immobilization in liquid culture by CD43- or CD44-antibody coating

Loeffler, Dirk ; Wang, Weijia ; Hopf, Alois ; Hilsenbeck, Oliver ; Bourgine, Paul LU orcid ; Rudolf, Fabian ; Martin, Ivan and Schroeder, Timm (2018) In Blood 131(13). p.1425-1429
Abstract
Keeping track of individual cell identifications is imperative to the study of dynamic single-cell behavior over time. Highly motile hematopoietic stem and progenitor cells (HSPCs) migrate quickly and do not adhere, and thus must be imaged very frequently to keep cell identifications. Even worse, they are also flushed away during medium exchange. To overcome these limitations, we tested antibody coating for reducing HSPC motility in vitro. Anti-CD43– and anti-CD44–antibody coating reduced the cell motility of mouse and human HSPCs in a concentration-dependent manner. This enables 2-dimensional (2D) colony formation without cell mixing in liquid cultures, massively increases time-lapse imaging throughput, and also maintains cell positions... (More)
Keeping track of individual cell identifications is imperative to the study of dynamic single-cell behavior over time. Highly motile hematopoietic stem and progenitor cells (HSPCs) migrate quickly and do not adhere, and thus must be imaged very frequently to keep cell identifications. Even worse, they are also flushed away during medium exchange. To overcome these limitations, we tested antibody coating for reducing HSPC motility in vitro. Anti-CD43– and anti-CD44–antibody coating reduced the cell motility of mouse and human HSPCs in a concentration-dependent manner. This enables 2-dimensional (2D) colony formation without cell mixing in liquid cultures, massively increases time-lapse imaging throughput, and also maintains cell positions during media exchange. Anti-CD43 but not anti-CD44 coating reduces mouse HSPC proliferation with increasing concentrations. No relevant effects on cell survival or myeloid and megakaryocyte differentiation of hematopoietic stem cells and multipotent progenitors 1-5 were detected. Human umbilical cord hematopoietic CD34+ cell survival, proliferation, and differentiation were not affected by either coating. This approach both massively simplifies and accelerates continuous analysis of suspension cells, and enables the study of their behavior in dynamic rather than static culture conditions over time. (Less)
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author
; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
in
Blood
volume
131
issue
13
pages
5 pages
publisher
American Society of Hematology
external identifiers
  • scopus:85047641061
ISSN
1528-0020
DOI
10.1182/blood-2017-07-794131
language
English
LU publication?
no
id
4c361fa4-decc-42f6-af71-ea1db9077c0a
date added to LUP
2022-02-09 19:01:11
date last changed
2022-04-20 04:50:53
@article{4c361fa4-decc-42f6-af71-ea1db9077c0a,
  abstract     = {{Keeping track of individual cell identifications is imperative to the study of dynamic single-cell behavior over time. Highly motile hematopoietic stem and progenitor cells (HSPCs) migrate quickly and do not adhere, and thus must be imaged very frequently to keep cell identifications. Even worse, they are also flushed away during medium exchange. To overcome these limitations, we tested antibody coating for reducing HSPC motility in vitro. Anti-CD43– and anti-CD44–antibody coating reduced the cell motility of mouse and human HSPCs in a concentration-dependent manner. This enables 2-dimensional (2D) colony formation without cell mixing in liquid cultures, massively increases time-lapse imaging throughput, and also maintains cell positions during media exchange. Anti-CD43 but not anti-CD44 coating reduces mouse HSPC proliferation with increasing concentrations. No relevant effects on cell survival or myeloid and megakaryocyte differentiation of hematopoietic stem cells and multipotent progenitors 1-5 were detected. Human umbilical cord hematopoietic CD34+ cell survival, proliferation, and differentiation were not affected by either coating. This approach both massively simplifies and accelerates continuous analysis of suspension cells, and enables the study of their behavior in dynamic rather than static culture conditions over time.}},
  author       = {{Loeffler, Dirk and Wang, Weijia and Hopf, Alois and Hilsenbeck, Oliver and Bourgine, Paul and Rudolf, Fabian and Martin, Ivan and Schroeder, Timm}},
  issn         = {{1528-0020}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{13}},
  pages        = {{1425--1429}},
  publisher    = {{American Society of Hematology}},
  series       = {{Blood}},
  title        = {{Mouse and human HSPC immobilization in liquid culture by CD43- or CD44-antibody coating}},
  url          = {{http://dx.doi.org/10.1182/blood-2017-07-794131}},
  doi          = {{10.1182/blood-2017-07-794131}},
  volume       = {{131}},
  year         = {{2018}},
}