Quantitative activation suppression assay to evaluate human bone marrow-derived mesenchymal stromal cell potency

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• 作者：Bahey Salem¹ ; ³ ; ^&lowast ; ; Samantha Miner¹ ; ^&lowast ; ; Nancy F. Hensel¹ ; Minoo Battiwalla¹ ; Keyvan Keyvanfar¹ ; David F. Stroncek² ; Adrian P. Gee³ ; Patrick J. Hanley⁴ ; Catherine M. Bollard⁴ ; Sawa Ito¹ ; ^&lowast ; ; ^{itos2@mail.nih.gov" class="auth_mail" title="E-mail the corresponding author} ; A. John Barrett¹ ; ^&lowast ;
• 关键词：immunosuppression ; suppression potency assay ; mesenchymal stromal cells ; K299
• 刊名：Cytotherapy
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：17
• 期：12
• 页码：1675-1686
• 全文大小：1781 K

文摘

With the increasing use of cell therapies involving immune modulatory cells, there is a need for a simple standardized method to evaluate and compare the suppressive potency of different cell products. We used the Karpas 299 (K299) cell line as the reference suppressor cell to develop a standardized suppression assay to quantify the immune-modulatory capacity of bone marrow–derived mesenchymal stromal cells (BM-MSCs).

Methods

Healthy donor CD4 T cells were co-cultured with the K299 cell line or with third-party BM-MSCs. After stimulation with anti-CD3/CD28 beads, CD154 activation and proliferation of CD4 T cells were measured to calculate suppression.

Results

The K299 cell line reproducibly suppressed both the activation and proliferation of healthy donor CD4 T cells in a dose-dependent manner. A rapid (16-h) assay that was based on activation-suppression was selected for development. In replicate testing, there was an inherent variability of suppression of 11% coefficient of variation between different responder T cells. Suppression by BM-MSCs on different responders correlated with suppression by K299. We therefore used K299 suppression as the reference to define suppression potency of BM-MSCs in K299 Suppression Units. We found that inter-donor variability, passage number, method of manufacture and exposure of BM-MSCs to steroids or interferon-γ all affected BM-MSC potency of suppression.

Conclusions

This method provides a platform for standardizing suppressor function to facilitate comparisons between laboratories and for use as a cell product release assay.