使用多种效价测定法评估人基质细胞。

Use of multiple potency assays to evaluate human mesenchymal stromal cells.

机构信息

From the Blood and Coagulation Research Department (B.A.C., M.C.H., C.P.D., I.A., C.C., C.S., S.L., L.G., K.J., R.M., A.P.C., J.A.B.), US Army Institute of Surgical Research, JBSA Fort Sam Houston, Houston; Department of Molecular Medicine (B.A.C.), UT Health San Antonio; Joint Interdisciplinary Program in Biomedical Engineering Program (A.P.C., J.A.B.), UT San Antonio and UT Health San Antonio; Department of Surgery (A.P.C.), UT Health San Antonio, San Antonio; and Institute of Biomedical Studies (J.A.B.), Baylor University, Waco, Texas.

出版信息

J Trauma Acute Care Surg. 2020 Aug;89(2S Suppl 2):S109-S117. doi: 10.1097/TA.0000000000002743.

DOI:10.1097/TA.0000000000002743

PMID:32744836

Abstract

BACKGROUND

There is broad interest in the use of cell therapies and cell products for treatment of a variety of diseases and problems. Of interest to the military, cellular therapies have the potential to confer tremendous benefit for treatment of both acute and chronic injuries. Although many different cell therapy products are currently under investigation, mesenchymal stromal cells (MSCs) are good candidates, based on their ability to respond to inflammation, limit vascular permeability, and modulate immune responses to injury. Although a large number of clinical trials utilize MSCs or their products, there is no firm consensus defining the characteristics and activities of a good MSC product. Here, we test multiple human MSCs in several assays designed to test potency, to determine if functionally relevant differences between MSCs can be defined using in vitro assays, allowing identification of superior MSC products for preclinical or clinical testing.

METHODS

Human MSCs derived from several tissue sources (adipose, bone marrow, umbilical cord) were evaluated for their ability to respond to inflammatory signaling by upregulating indoleamine-2,3-dioxygenase and TSG6, suppress lymphocyte proliferation, alter the polarization of macrophages, and affect tube formation by endothelial cells.

RESULTS

All MSCs tested displayed activity in the functional assays utilized, but differences in potency were observed in each assay.

CONCLUSION

The indoleamine-2,3-dioxygenase enzyme activity assay represents a simple way to screen multiple samples. The mixed lymphocyte reaction and monocyte assays used to test interactions between MSCs and immune cells are more involved but give direct information on immunomodulation potential. The endothelial cell tube formation assay is relatively simple to perform but a large number of images must be generated and analyzed. However, it tests a functional activity other than immunomodulation and, therefore, adds another facet to MSC evaluation.

摘要

背景

细胞疗法和细胞产品在治疗各种疾病和问题方面的应用引起了广泛关注。对于军队来说，细胞疗法有可能为治疗急性和慢性损伤带来巨大益处。尽管目前有许多不同的细胞治疗产品正在研究中，但间充质基质细胞（MSCs）是很好的候选者，因为它们能够对炎症做出反应，限制血管通透性，并调节对损伤的免疫反应。尽管大量的临床试验利用 MSCs 或其产品，但对于一个好的 MSC 产品的特征和活性还没有明确的共识。在这里，我们在多个旨在测试效力的测定中测试了多种人 MSCs，以确定是否可以使用体外测定来定义 MSCs 之间的功能相关差异，从而确定用于临床前或临床测试的更好的 MSC 产品。

方法

从几种组织来源（脂肪、骨髓、脐带）中提取的人 MSCs 被评估其通过上调吲哚胺 2,3-双加氧酶和 TSG6 来响应炎症信号的能力，抑制淋巴细胞增殖，改变巨噬细胞的极化，并影响内皮细胞的管形成。

结果

所有测试的 MSCs 在使用的功能测定中都显示出活性，但在每个测定中都观察到效力的差异。

结论

色氨酸 2,3-双加氧酶酶活性测定是筛选多个样本的简单方法。混合淋巴细胞反应和单核细胞测定用于测试 MSCs 与免疫细胞之间的相互作用，虽然更复杂，但直接提供了免疫调节潜力的信息。内皮细胞管形成测定相对简单，但必须生成和分析大量的图像。然而，它测试了除免疫调节以外的另一种功能活性，因此为 MSC 评估增加了另一个方面。