Identification of fetal liver stroma in spectral cytometry using the parameter autofluorescence†

Autor: Francisca Soares-da-Silva, Marie-Pierre Mailhé, Ait-Mansour C, Marcia Peixoto, Ana Cumano, Sophie Novault, Sandrine Schmutz
Přispěvatelé: Novault, Sophie, Appel à projets générique - Deux progéniteurs hématopoïétiques différents établissent le compartiment de lymphocytes T: tester un nouveau paradigme du développement T. - - Twothyme2014 - ANR-14-CE11-0022 - Appel à projets générique - VALID, Développement des cellules lymphoïdes embryonnaires qui organisent l'architecture du thymus - - DELSTAR2021 - ANR-21-CE15-0028 - AAPG2021 - VALID, Blanc - A comprehensive approach to the study of epigenetic regulation of gene expression during development by the C. elegans HP1 proteins - - EpiDev2007 - ANR-07-BLAN-0081 - BLANC - VALID, Laboratoires d'excellence - Stem Cells in Regenerative Biology and Medicine - - REVIVE2010 - ANR-10-LABX-0073 - LABX - VALID, Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Universidade do Porto = University of Porto, Cytométrie (plateforme) - Flow Cytometry (platform), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Sony Corporation, This work was financed by the Institut Pasteur, INSERM, ANR (grant Twothyme, grant DELSTAR and grant EPI-DEV), REVIVE Future Investment Program and Pasteur-Weizmann Foundation through grants to A.C. This work was financed by Portuguese funds through FCT/MCTES in the framework of the project PTDC/MED-OUT/32656/2017 (POCI-01-0145-FEDER-032656). F.S.S is funded by the REVIVE Future investments post-doctoral grant (Investissement d'Avenir, ANR-10-LABX-73). MP is funded by FCT grant SFRH/BD/143605/2019, We thank all members of the flow cytometry core facility, the members of the Institut Pasteur laboratory, Antonio Bandeira, Paulo Vieira, Rachel Golub and Pablo Pereira., ANR-14-CE11-0022,Twothyme,Deux progéniteurs hématopoïétiques différents établissent le compartiment de lymphocytes T: tester un nouveau paradigme du développement T.(2014), ANR-21-CE15-0028,DELSTAR,Développement des cellules lymphoïdes embryonnaires qui organisent l'architecture du thymus(2021), ANR-07-BLAN-0081,EpiDev,A comprehensive approach to the study of epigenetic regulation of gene expression during development by the C. elegans HP1 proteins(2007), ANR-10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010)
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Cytometry Part A
Cytometry Part A, 2022, ⟨10.1002/cyto.a.24567⟩
ISSN: 1552-4922
1552-4930
Popis: The fetal liver is the main hematopoietic organ during embryonic development. The fetal liver is also the unique anatomical site where hematopoietic stem cells expand before colonizing the bone marrow, where they ensure life-long blood cell production and become mostly resting. The identification of the different cell types that comprise the hematopoietic stroma in the fetal liver is essential to understand the signals required for the expansion and differentiation of the hematopoietic stem cells. We used a panel of monoclonal antibodies to identify fetal liver stromal cells in a 5-laser equipped spectral flow cytometry analyzer. The “Autofluorescence Finder” of SONY ID7000 software identified two distinct autofluorescence emission spectra. Using autofluorescence as a fluorescence parameter we could assign the two autofluorescent signals to three distinct cell types and identified surface markers that characterize these populations. We found that one autofluorescent population corresponds to hepatoblasts and cholangiocytes whereas the other expresses mesenchymal transcripts and was identified as stellate cells. Importantly, after birth, autofluorescence becomes the unique identifying property of hepatoblasts because mature cholangiocytes are no longer autofluorescent.These results show that autofluorescence used as a parameter in spectral flow cytometry is a useful tool to identify new cell subsets that are difficult to analyze in conventional flow cytometry.
Databáze: OpenAIRE
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