Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties

Abstract Background Umbilical Cord-derived Mesenchymal Stromal Cells (UC-MSCs) display high immunoregulatory properties, offering new perspectives to treat severe immune and inflammatory diseases. However, the heterogeneity of their biological properties remains a challenge to predict clinical respo...

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Main Authors: Miryam Mebarki, Coralie Moine-Picard, Romain Enjaume-Rauch, Antoine Laurent-Puig, Annaelle Suissa, Valentine Feyants, Jérôme Larghero, Audrey Cras
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-025-04361-y
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author Miryam Mebarki
Coralie Moine-Picard
Romain Enjaume-Rauch
Antoine Laurent-Puig
Annaelle Suissa
Valentine Feyants
Jérôme Larghero
Audrey Cras
author_facet Miryam Mebarki
Coralie Moine-Picard
Romain Enjaume-Rauch
Antoine Laurent-Puig
Annaelle Suissa
Valentine Feyants
Jérôme Larghero
Audrey Cras
author_sort Miryam Mebarki
collection DOAJ
description Abstract Background Umbilical Cord-derived Mesenchymal Stromal Cells (UC-MSCs) display high immunoregulatory properties, offering new perspectives to treat severe immune and inflammatory diseases. However, the heterogeneity of their biological properties remains a challenge to predict clinical response. The aim of our study is to evaluate a strategy based on the constitution of a pool of several pre-selected donors to reduce the biological variability of UC-MSCs and improve their immunomodulatory properties. Methods Umbilical cords were collected from 10 healthy donors. Isolated UC-MSCs were characterized in the basal state and after a pro-inflammatory priming in vitro by interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα). Proliferation, immunophenotype, the expression of activation markers and the inhibition of T cell proliferation in vitro were assessed in UC-MSCs from selected single donors and from pools. Results Our study highlights the donor-dependent heterogeneity of UC-MSCs immunomodulatory functions. Based on their ability to suppress T-cell proliferation in vitro, we classified donors into three profiles: high, medium and low. Preparation of pools containing UC-MSCs derived from each profile in a 1:1:1 ratio reduced the donor-dependent variability and, most importantly, improved the lowest immunomodulatory functions. After priming with pro-inflammatory cytokines, the inhibition of T-cell expansion by the pooled UC-MSCs was significantly higher than the low donor and the theoretical mean of individual donors, and was associated with increased expression of the key immunoregulatory proteins. Interestingly, the pool did not induce a cumulative immunogenic effect: expression of HLA or costimulatory molecules between the high donor and the pool were similar. Finally, pooling UC-MSCs derived from high and low donors in a 1:2 ratio was sufficient to enhance the lowest immunomodulatory properties. Conclusion Overall, our results demonstrate that pooled UC-MSCs with selected high donor offers a new strategy to optimize the immunomodulatory functions of allogeneic UC-MSC-based medicinal products. Graphical Abstract
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spelling doaj-art-d333f6c84cf745b29f226db4b4a11b822025-08-20T03:48:19ZengBMCStem Cell Research & Therapy1757-65122025-05-0116111610.1186/s13287-025-04361-yPooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory propertiesMiryam Mebarki0Coralie Moine-Picard1Romain Enjaume-Rauch2Antoine Laurent-Puig3Annaelle Suissa4Valentine Feyants5Jérôme Larghero6Audrey Cras7AP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAP-HP, Unité de Thérapie Cellulaire, INSERM Centre d’Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-LouisAbstract Background Umbilical Cord-derived Mesenchymal Stromal Cells (UC-MSCs) display high immunoregulatory properties, offering new perspectives to treat severe immune and inflammatory diseases. However, the heterogeneity of their biological properties remains a challenge to predict clinical response. The aim of our study is to evaluate a strategy based on the constitution of a pool of several pre-selected donors to reduce the biological variability of UC-MSCs and improve their immunomodulatory properties. Methods Umbilical cords were collected from 10 healthy donors. Isolated UC-MSCs were characterized in the basal state and after a pro-inflammatory priming in vitro by interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα). Proliferation, immunophenotype, the expression of activation markers and the inhibition of T cell proliferation in vitro were assessed in UC-MSCs from selected single donors and from pools. Results Our study highlights the donor-dependent heterogeneity of UC-MSCs immunomodulatory functions. Based on their ability to suppress T-cell proliferation in vitro, we classified donors into three profiles: high, medium and low. Preparation of pools containing UC-MSCs derived from each profile in a 1:1:1 ratio reduced the donor-dependent variability and, most importantly, improved the lowest immunomodulatory functions. After priming with pro-inflammatory cytokines, the inhibition of T-cell expansion by the pooled UC-MSCs was significantly higher than the low donor and the theoretical mean of individual donors, and was associated with increased expression of the key immunoregulatory proteins. Interestingly, the pool did not induce a cumulative immunogenic effect: expression of HLA or costimulatory molecules between the high donor and the pool were similar. Finally, pooling UC-MSCs derived from high and low donors in a 1:2 ratio was sufficient to enhance the lowest immunomodulatory properties. Conclusion Overall, our results demonstrate that pooled UC-MSCs with selected high donor offers a new strategy to optimize the immunomodulatory functions of allogeneic UC-MSC-based medicinal products. Graphical Abstracthttps://doi.org/10.1186/s13287-025-04361-yHuman umbilical cordWharton’s jellyMesenchymal stromal cellsImmunomodulationDonor heterogeneityDonor variability
spellingShingle Miryam Mebarki
Coralie Moine-Picard
Romain Enjaume-Rauch
Antoine Laurent-Puig
Annaelle Suissa
Valentine Feyants
Jérôme Larghero
Audrey Cras
Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
Stem Cell Research & Therapy
Human umbilical cord
Wharton’s jelly
Mesenchymal stromal cells
Immunomodulation
Donor heterogeneity
Donor variability
title Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
title_full Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
title_fullStr Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
title_full_unstemmed Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
title_short Pooling umbilical cord-mesenchymal stromal cells derived from selected multiple donors reduces donor-dependent variability and improves their immunomodulatory properties
title_sort pooling umbilical cord mesenchymal stromal cells derived from selected multiple donors reduces donor dependent variability and improves their immunomodulatory properties
topic Human umbilical cord
Wharton’s jelly
Mesenchymal stromal cells
Immunomodulation
Donor heterogeneity
Donor variability
url https://doi.org/10.1186/s13287-025-04361-y
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