预处理间充质基质细胞球以分泌免疫调节旁分泌因子。

Pre-conditioning mesenchymal stromal cell spheroids for immunomodulatory paracrine factor secretion.

机构信息

The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University, Atlanta, Georgia, USA.

The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University, Atlanta, Georgia, USA; The Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA.

出版信息

Cytotherapy. 2014 Mar;16(3):331-45. doi: 10.1016/j.jcyt.2013.09.004. Epub 2013 Nov 9.

DOI:10.1016/j.jcyt.2013.09.004

PMID:24219905

Abstract

BACKGROUND AIMS

Mesenchymal stromal cells (MSCs) exhibit the inherent potential to regulate multiple signaling pathways and cell types that contribute to the pathogenesis of inflammatory and immune diseases. However, more recent studies have suggested that the secretion of immunomodulatory factors by MSCs can be enhanced by three-dimensional aggregation or pro-inflammatory cytokine treatment.

METHODS

Human MSC spheroids were formed by forced aggregation into agarose micro-wells and subsequently cultured in either minimal essential medium alpha supplemented with fetal bovine serum or serum-free, defined MesenCult-XF medium (STEMCELL Technologies, Vancouver, Canada). A subset of the spheroids were treated with pro-inflammatory cytokines interferon (IFN)-γ or tumor necrosis factor (TNF)-α or both for 4 days. Immunomodulatory factor (prostaglandin E2, indoleamine 2,3-dioxygenase, transforming growth factor-β1 and interleukin-6) secretion was quantified after 4 days of culture, and the immunomodulatory activity of MSCs was assessed by quantifying activated macrophage expression of TNF-α after trans-well co-culture.

RESULTS

Culturing human MSCs as three-dimensional aggregates increased secretion of immunomodulatory paracrine factors, which was enhanced further by treatment with IFN-γ and TNF-α, demonstrating that these parameters can synergistically enhance endogenous human MSC immunomodulatory properties. However, immunomodulatory factor secretion was found to be highly dependent on the composition of cell culture medium. Human MSCs cultured in MesenCult-XF medium displayed significantly less expression of prostaglandin E2, indoleamine 2,3-dioxygenase, transforming growth factor-β1 and interleukin-6 compared with human MSCs cultured in medium supplemented with fetal bovine serum. Finally, pre-conditioning of human MSC spheroids with IFN-γ and TNF-α resulted in greater immunomodulatory activity in a macrophage co-culture assay.

CONCLUSIONS

Altogether, engineering the environment of human MSCs to develop pre-conditioning strategies for enhancing human MSC immunomodulation may be a simple approach for improving MSC-based therapies for the treatment of inflammatory and immune diseases.

摘要

背景目的

间充质基质细胞（MSCs）表现出固有潜力，可调节多种信号通路和细胞类型，从而促进炎症和免疫性疾病的发病机制。然而，最近的研究表明，MSCs 免疫调节因子的分泌可以通过三维聚集或促炎细胞因子处理来增强。

方法

通过强制聚集到琼脂糖微井中来形成人 MSC 球体，然后在补充有胎牛血清的最低必需培养基 alpha 或无血清、定义明确的 Mesencult-XF 培养基（STEMCELL Technologies，温哥华，加拿大）中进行培养。一部分球体用促炎细胞因子干扰素（IFN）-γ或肿瘤坏死因子（TNF）-α或两者处理 4 天。培养 4 天后定量测定免疫调节因子（前列腺素 E2、吲哚胺 2,3-双加氧酶、转化生长因子-β1 和白细胞介素-6）的分泌，并通过定量穿过培养物共培养的活化巨噬细胞 TNF-α的表达来评估 MSC 的免疫调节活性。

结果

将人 MSC 培养为三维聚集体增加了免疫调节旁分泌因子的分泌，用 IFN-γ和 TNF-α处理进一步增强了这种分泌，表明这些参数可以协同增强内源性人 MSC 的免疫调节特性。然而，免疫调节因子的分泌高度依赖于细胞培养基的组成。与补充有胎牛血清的培养基相比，在 Mesencult-XF 培养基中培养的人 MSC 显示出明显较少的前列腺素 E2、吲哚胺 2,3-双加氧酶、转化生长因子-β1 和白细胞介素-6 的表达。最后，用 IFN-γ和 TNF-α预处理人 MSC 球体，在巨噬细胞共培养测定中导致更大的免疫调节活性。