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Identification and characterisation of maternal perivascular SUSD2+ placental mesenchymal stem/stromal cells

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Abstract

Mesenchymal stem cells (MSCs) that meet the International Society for Cellular Therapy (ISCT) criteria are obtained from placental tissue by plastic adherence. Historically, no known single marker was available for isolating placental MSCs (pMSCs) from the decidua basalis. As the decidua basalis is derived from the regenerative endometrium, we hypothesised that SUSD2, an endometrial perivascular MSC marker, would purify maternal perivascular pMSC. Perivascular pMSCs were isolated from the maternal placenta using SUSD2 magnetic bead sorting and assessed for the colony-forming unit-fibroblasts (CFU-F), surface markers, and in vitro differentiation into mesodermal lineages. Multi-colour immunofluorescence was used to colocalise SUSD2 and α-SMA, a perivascular marker in the decidua basalis. Placental stromal cell suspensions comprised 5.1%SUSD2+ cells. SUSD2 magnetic bead sorting of the placental stromal cells increased their purity approximately two-fold. SUSD2+ pMSCs displayed greater CFU-F activity than SUSD2− stromal fibroblasts (pSFs). However, both SUSD2+ pMSC and SUSD2− pSF underwent mesodermal differentiation in vitro, and both expressed the ISCT surface markers. Higher percentages of cultured SUSD2+ pMSCs expressed the perivascular markers CD146, CD140b, and SUSD2 than SUSD2− pSFs. These findings suggest that SUSD2 is a single marker that enriches maternal pMSCs, suggesting they may originate from eMSC. Placental decidua basalis can be used as an alternative source of MSC for clinical translation in situations where there is no access to endometrial tissue.

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Acknowledgements

The authors acknowledge Grace Heo and Jenny Ryan for collection of human tissues; Angela Vais, Monash Histology Platform, for assistance with histology and specific staining; and Dr Sarah Creed and Dr Giulia Ballerin, Monash Micro Imaging, for their excellent technical support in confocal microscope imaging.

Funding

This work was supported by the National Health and Medical Research Council (NHMRC) of Australia Project Grants (No. 1159677 to CEG, SM, and JAW), Senior Research Fellowship (No. 1042298 to CEG), and Investigator Grant (No. 1173882 to CEG) and Science and Industrial Endowment Fund (PD16-122 to CEG, JW, and SM), Monash Graduate Scholarship (MK), Monash International Tuition Scholarship (MK), and the Victorian Government’s Operational Infrastructure Support Program.

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  1. Fiona L. Cousins and Caroline E. Gargett are equal senior authors

Authors and Affiliations

  1. The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia

    Manijeh Khanmohammadi, Shayanti Mukherjee, Saeedeh Darzi, Kallyanashis Paul, Jerome A. Werkmeister, Fiona L. Cousins & Caroline E. Gargett

  2. Obstetrics and Gynaecology, Monash University, Clayton, VIC, Australia

    Manijeh Khanmohammadi, Shayanti Mukherjee, Saeedeh Darzi, Kallyanashis Paul, Jerome A. Werkmeister, Fiona L. Cousins & Caroline E. Gargett

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  1. Manijeh Khanmohammadi
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Correspondence to Caroline E. Gargett.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the National Health and Medical Research Council of Australia Guidelines as approved by Monash Health Human Research Ethics Committee (Ethics Number: 10103B) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All participants gave written informed consent. This article does not contain any studies with animals performed by any of the authors.

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Khanmohammadi, M., Mukherjee, S., Darzi, S. et al. Identification and characterisation of maternal perivascular SUSD2+ placental mesenchymal stem/stromal cells. Cell Tissue Res 385, 803–815 (2021). https://doi.org/10.1007/s00441-021-03453-4

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