---
_id: '11478'
abstract:
- lang: eng
  text: Cerebral organoids differentiated from human-induced pluripotent stem cells
    (hiPSC) provide a unique opportunity to investigate brain development. However,
    organoids usually lack microglia, brain-resident immune cells, which are present
    in the early embryonic brain and participate in neuronal circuit development.
    Here, we find IBA1+ microglia-like cells alongside retinal cups between week 3
    and 4 in 2.5D culture with an unguided retinal organoid differentiation protocol.
    Microglia do not infiltrate the neuroectoderm and instead enrich within non-pigmented,
    3D-cystic compartments that develop in parallel to the 3D-retinal organoids. When
    we guide the retinal organoid differentiation with low-dosed BMP4, we prevent
    cup development and enhance microglia and 3D-cysts formation. Mass spectrometry
    identifies these 3D-cysts to express mesenchymal and epithelial markers. We confirmed
    this microglia-preferred environment also within the unguided protocol, providing
    insight into microglial behavior and migration and offer a model to study how
    they enter and distribute within the human brain.
acknowledged_ssus:
- _id: Bio
- _id: LifeSc
acknowledgement: We thank the scientific service units at ISTA, specifically the lab
  support facility and imaging & optics facility for their support; Nicolas Armel
  for performing the Mass Spectrometry. We thank Alexandra Lang and Tanja Peilnsteiner
  for their help in human brain tissue collection, Rouven Schulz for his insights
  into the functional assays We thank all members of the Siegert group for constant
  feedback on the project and Margaret Maes, Rouven Schulz, and Marco Benevento for
  feedback on the manuscript. This project has received funding from the European
  Research Council (ERC) under the European Union’s Horizon 2020 research and innovation
  program (grant No. 715571 to S.S.) and from the Gesellschaft für Forschungsförderung
  Niederösterreich (grant No. Sc19-017 to V.H.).
article_number: '104580'
article_processing_charge: Yes
article_type: original
author:
- first_name: Katarina
  full_name: Bartalska, Katarina
  id: 4D883232-F248-11E8-B48F-1D18A9856A87
  last_name: Bartalska
- first_name: Verena
  full_name: Hübschmann, Verena
  id: 32B7C918-F248-11E8-B48F-1D18A9856A87
  last_name: Hübschmann
- first_name: Medina
  full_name: Korkut, Medina
  id: 4B51CE74-F248-11E8-B48F-1D18A9856A87
  last_name: Korkut
  orcid: 0000-0003-4309-2251
- first_name: Ryan J
  full_name: Cubero, Ryan J
  id: 850B2E12-9CD4-11E9-837F-E719E6697425
  last_name: Cubero
  orcid: 0000-0003-0002-1867
- first_name: Alessandro
  full_name: Venturino, Alessandro
  id: 41CB84B2-F248-11E8-B48F-1D18A9856A87
  last_name: Venturino
  orcid: 0000-0003-2356-9403
- first_name: Karl
  full_name: Rössler, Karl
  last_name: Rössler
- first_name: Thomas
  full_name: Czech, Thomas
  last_name: Czech
- first_name: Sandra
  full_name: Siegert, Sandra
  id: 36ACD32E-F248-11E8-B48F-1D18A9856A87
  last_name: Siegert
  orcid: 0000-0001-8635-0877
citation:
  ama: Bartalska K, Hübschmann V, Korkut M, et al. A systematic characterization of
    microglia-like cell occurrence during retinal organoid differentiation. <i>iScience</i>.
    2022;25(7). doi:<a href="https://doi.org/10.1016/j.isci.2022.104580">10.1016/j.isci.2022.104580</a>
  apa: Bartalska, K., Hübschmann, V., Korkut, M., Cubero, R. J., Venturino, A., Rössler,
    K., … Siegert, S. (2022). A systematic characterization of microglia-like cell
    occurrence during retinal organoid differentiation. <i>IScience</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.isci.2022.104580">https://doi.org/10.1016/j.isci.2022.104580</a>
  chicago: Bartalska, Katarina, Verena Hübschmann, Medina Korkut, Ryan J Cubero, Alessandro
    Venturino, Karl Rössler, Thomas Czech, and Sandra Siegert. “A Systematic Characterization
    of Microglia-like Cell Occurrence during Retinal Organoid Differentiation.” <i>IScience</i>.
    Elsevier, 2022. <a href="https://doi.org/10.1016/j.isci.2022.104580">https://doi.org/10.1016/j.isci.2022.104580</a>.
  ieee: K. Bartalska <i>et al.</i>, “A systematic characterization of microglia-like
    cell occurrence during retinal organoid differentiation,” <i>iScience</i>, vol.
    25, no. 7. Elsevier, 2022.
  ista: Bartalska K, Hübschmann V, Korkut M, Cubero RJ, Venturino A, Rössler K, Czech
    T, Siegert S. 2022. A systematic characterization of microglia-like cell occurrence
    during retinal organoid differentiation. iScience. 25(7), 104580.
  mla: Bartalska, Katarina, et al. “A Systematic Characterization of Microglia-like
    Cell Occurrence during Retinal Organoid Differentiation.” <i>IScience</i>, vol.
    25, no. 7, 104580, Elsevier, 2022, doi:<a href="https://doi.org/10.1016/j.isci.2022.104580">10.1016/j.isci.2022.104580</a>.
  short: K. Bartalska, V. Hübschmann, M. Korkut, R.J. Cubero, A. Venturino, K. Rössler,
    T. Czech, S. Siegert, IScience 25 (2022).
date_created: 2022-07-03T22:01:33Z
date_published: 2022-07-15T00:00:00Z
date_updated: 2023-11-02T12:21:33Z
day: '15'
ddc:
- '610'
department:
- _id: SaSi
doi: 10.1016/j.isci.2022.104580
ec_funded: 1
external_id:
  isi:
  - '000830428500005'
file:
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- iso: eng
month: '07'
oa: 1
oa_version: Published Version
project:
- _id: 25D4A630-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '715571'
  name: Microglia action towards neuronal circuit formation and function in health
    and disease
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  name: IST Austria Open Access Fund
- _id: 9B99D380-BA93-11EA-9121-9846C619BF3A
  grant_number: SC19-017
  name: How human microglia shape developing neurons during health and inflammation
publication: iScience
publication_identifier:
  eissn:
  - 2589-0042
publication_status: published
publisher: Elsevier
quality_controlled: '1'
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scopus_import: '1'
status: public
title: A systematic characterization of microglia-like cell occurrence during retinal
  organoid differentiation
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
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type: journal_article
user_id: 3E5EF7F0-F248-11E8-B48F-1D18A9856A87
volume: 25
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...
---
_id: '12117'
abstract:
- lang: eng
  text: "To understand how potential gene manipulations affect in vitro microglia,
    we provide a set of short protocols to evaluate microglia identity and function.
    We detail steps for immunostaining to determine microglia identity. We describe
    three functional assays for microglia: phagocytosis, calcium response following
    ATP stimulation, and cytokine expression upon inflammatory stimuli. We apply these
    protocols to human induced-pluripotent-stem-cell (hiPSC)-derived microglia, but
    they can be also applied to other in vitro microglial models including primary
    mouse microglia.\r\nFor complete details on the use and execution of this protocol,
    please refer to Bartalska et al. (2022).1"
acknowledged_ssus:
- _id: Bio
acknowledgement: This project has received funding from the European Research Council
  (ERC) under the European Union’s Horizon 2020 research and innovation program (grant
  No. 715571 to S.S.) and from the Gesellschaft für Forschungsförderung Niederösterreich
  (grant No. Sc19-017 to V.H.). We thank Rouven Schulz and Alessandro Venturino for
  their insights into functional assays and data analysis, Verena Seiboth for insights
  into necessary institutional permission, and ISTA imaging & optics facility (IOF)
  especially Bernhard Hochreiter for their support.
article_number: '101866'
article_processing_charge: No
article_type: letter_note
author:
- first_name: Verena
  full_name: Hübschmann, Verena
  id: 32B7C918-F248-11E8-B48F-1D18A9856A87
  last_name: Hübschmann
- first_name: Medina
  full_name: Korkut, Medina
  id: 4B51CE74-F248-11E8-B48F-1D18A9856A87
  last_name: Korkut
  orcid: 0000-0003-4309-2251
- first_name: Sandra
  full_name: Siegert, Sandra
  id: 36ACD32E-F248-11E8-B48F-1D18A9856A87
  last_name: Siegert
  orcid: 0000-0001-8635-0877
citation:
  ama: Hübschmann V, Korkut M, Siegert S. Assessing human iPSC-derived microglia identity
    and function by immunostaining, phagocytosis, calcium activity, and inflammation
    assay. <i>STAR Protocols</i>. 2022;3(4). doi:<a href="https://doi.org/10.1016/j.xpro.2022.101866">10.1016/j.xpro.2022.101866</a>
  apa: Hübschmann, V., Korkut, M., &#38; Siegert, S. (2022). Assessing human iPSC-derived
    microglia identity and function by immunostaining, phagocytosis, calcium activity,
    and inflammation assay. <i>STAR Protocols</i>. Elsevier. <a href="https://doi.org/10.1016/j.xpro.2022.101866">https://doi.org/10.1016/j.xpro.2022.101866</a>
  chicago: Hübschmann, Verena, Medina Korkut, and Sandra Siegert. “Assessing Human
    IPSC-Derived Microglia Identity and Function by Immunostaining, Phagocytosis,
    Calcium Activity, and Inflammation Assay.” <i>STAR Protocols</i>. Elsevier, 2022.
    <a href="https://doi.org/10.1016/j.xpro.2022.101866">https://doi.org/10.1016/j.xpro.2022.101866</a>.
  ieee: V. Hübschmann, M. Korkut, and S. Siegert, “Assessing human iPSC-derived microglia
    identity and function by immunostaining, phagocytosis, calcium activity, and inflammation
    assay,” <i>STAR Protocols</i>, vol. 3, no. 4. Elsevier, 2022.
  ista: Hübschmann V, Korkut M, Siegert S. 2022. Assessing human iPSC-derived microglia
    identity and function by immunostaining, phagocytosis, calcium activity, and inflammation
    assay. STAR Protocols. 3(4), 101866.
  mla: Hübschmann, Verena, et al. “Assessing Human IPSC-Derived Microglia Identity
    and Function by Immunostaining, Phagocytosis, Calcium Activity, and Inflammation
    Assay.” <i>STAR Protocols</i>, vol. 3, no. 4, 101866, Elsevier, 2022, doi:<a href="https://doi.org/10.1016/j.xpro.2022.101866">10.1016/j.xpro.2022.101866</a>.
  short: V. Hübschmann, M. Korkut, S. Siegert, STAR Protocols 3 (2022).
date_created: 2023-01-12T11:56:38Z
date_published: 2022-12-16T00:00:00Z
date_updated: 2023-11-02T12:21:32Z
day: '16'
ddc:
- '570'
department:
- _id: SaSi
- _id: GradSch
doi: 10.1016/j.xpro.2022.101866
ec_funded: 1
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  date_created: 2023-01-23T09:50:51Z
  date_updated: 2023-01-23T09:50:51Z
  file_id: '12340'
  file_name: 2022_STARProtocols_Huebschmann.pdf
  file_size: 6251945
  relation: main_file
  success: 1
file_date_updated: 2023-01-23T09:50:51Z
has_accepted_license: '1'
intvolume: '         3'
issue: '4'
keyword:
- General Immunology and Microbiology
- General Biochemistry
- Genetics and Molecular Biology
- General Neuroscience
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
project:
- _id: 25D4A630-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '715571'
  name: Microglia action towards neuronal circuit formation and function in health
    and disease
- _id: 9B99D380-BA93-11EA-9121-9846C619BF3A
  grant_number: SC19-017
  name: How human microglia shape developing neurons during health and inflammation
publication: STAR Protocols
publication_identifier:
  issn:
  - 2666-1667
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  record:
  - id: '11478'
    relation: other
    status: public
scopus_import: '1'
status: public
title: Assessing human iPSC-derived microglia identity and function by immunostaining,
  phagocytosis, calcium activity, and inflammation assay
tmp:
  image: /images/cc_by_nc_nd.png
  legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
  name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
    (CC BY-NC-ND 4.0)
  short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 3
year: '2022'
...