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

被引：31

作者：

Salem, Bahey ^{[1
,3
,4
]}

Miner, Samantha ^{[1
]}

Hensel, Nancy F. ^{[1
]}

Battiwalla, Minoo ^{[1
]}

Keyvanfar, Keyvan ^{[1
]}

Stroncek, David F. ^{[2
]}

Gee, Adrian P. ^{[3
,4
]}

Hanley, Patrick J. ^{[5
,6
]}

Bollard, Catherine M. ^{[5
,6
]}

Ito, Sawa ^{[1
]}

Barrett, A. John ^{[1
]}

机构：

[1] NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA

[2] NIH, Dept Transfus Med, Bethesda, MD USA

[3] Houston Methodist Hosp, Texas Childrens Hosp, Ctr Cell & Gene Therapy, Houston, TX USA

[4] Baylor Coll Med, Houston, TX 77030 USA

[5] Childrens Natl Hlth Syst, Program Cell Enhancement & Technol Immunotherapy, Ctr Canc & Immunol Res, Sheikh Zayed Inst Pediat Surg Innovat, Washington, DC USA

[6] Childrens Natl Hlth Syst, Div Blood & Marrow Transplantat, Washington, DC USA

来源：

CYTOTHERAPY | 2015年 / 17卷 / 12期

基金：

美国国家卫生研究院;

关键词：

immunosuppression; suppression potency assay; mesenchymal stromal cells; K299; REGULATORY T-CELL; VERSUS-HOST-DISEASE; STEM-CELLS; MEMORY CELLS; EXPANSION; TRANSPLANTATION; KARPAS-299; PHENOTYPE; EFFECTOR; RECEPTOR;

D O I：

10.1016/j.jcyt.2015.08.008

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Background aims. 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, CD 154 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-gamma 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.

引用

页码：1675 / 1686

页数：12

共 31 条

[1] Surface phenotype and antigenic specificity of human interleukin 17-producing T helper memory cells [J].