Mebarki Miryam, Moine-Picard Coralie, Enjaume-Rauch Romain, Laurent-Puig Antoine, Suissa Annaelle, Feyants Valentine, Larghero Jérôme, Cras Audrey
AP-HP, Unité de Thérapie Cellulaire, INSERM Centre d'Investigation Clinique en Biothérapies CIC-BT, Hôpital Saint-Louis, 75010, Paris, France.
INSERM UMR1342, Université Paris Cité, 75010, Paris, France.
Stem Cell Res Ther. 2025 May 20;16(1):252. doi: 10.1186/s13287-025-04361-y.
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.
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.
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.
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.
脐带间充质基质细胞（UC-MSCs）具有高度免疫调节特性，为治疗严重免疫和炎症性疾病提供了新的视角。然而，其生物学特性的异质性仍然是预测临床反应的一个挑战。我们研究的目的是评估一种基于构建多个预先选定供体库的策略，以降低UC-MSCs的生物学变异性并改善其免疫调节特性。
从10名健康供体收集脐带。分离的UC-MSCs在基础状态下以及在体外经干扰素-γ（IFNγ）和肿瘤坏死因子-α（TNFα）进行促炎预处理后进行表征。对来自选定单一供体和供体库的UC-MSCs的增殖、免疫表型、激活标志物的表达以及体外T细胞增殖的抑制进行评估。
我们的研究突出了UC-MSCs免疫调节功能的供体依赖性异质性。基于它们在体外抑制T细胞增殖的能力，我们将供体分为三种类型：高、中、低。以1:1:1的比例制备包含来自每种类型的UC-MSCs的供体库降低了供体依赖性变异性，最重要的是，改善了最低的免疫调节功能。在用促炎细胞因子预处理后，联合UC-MSCs对T细胞扩增的抑制明显高于低供体以及单个供体的理论平均值，并且与关键免疫调节蛋白的表达增加相关。有趣的是，供体库没有诱导累积免疫原性效应：高供体和供体库之间HLA或共刺激分子的表达相似。最后，以1:2的比例混合来自高供体和低供体的UC-MSCs足以增强最低的免疫调节特性。
总体而言，我们的结果表明，将选定的高供体的UC-MSCs混合提供了一种优化基于同种异体UC-MSC的药品免疫调节功能的新策略。
