脂肪组织来源的干细胞在软骨分化后表现出免疫原性和免疫抑制特性。

Adipose tissue-derived stem cells show both immunogenic and immunosuppressive properties after chondrogenic differentiation.

机构信息

Department of Otorhinolaryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University of Wuerzburg, Wuerzburg, Germany.

出版信息

Cytotherapy. 2011 Mar;13(3):310-7. doi: 10.3109/14653249.2010.504769. Epub 2010 Aug 26.

DOI:10.3109/14653249.2010.504769

PMID:20795757

Abstract

BACKGROUND AIMS

The chondrogenic differentiation potential of mesenchymal stromal cells (MSC), as well as their immunosuppressive properties, have been studied extensively. So far, only a few studies have addressed the question of whether MSC still retain their immunosuppressive qualities after transdifferentiation. In particular, the expression of immunogenic markers, such as human leukocyte antigen (HLA)-DR, after differentiation has never been investigated.

METHODS

Chondrogenic transdifferentiation was induced in human adipose tissue-derived stem cell (ADSC) pellet cultures derived from 10 different patients, using 10 ng/mL transforming growth factor (TGF)-β3. Samples were harvested over a time-course of 28 days and analyzed by immunohistochemistry and reverse transcription (RT)-polymerase chain reaction (PCR). The cytokine levels in the supernatants of the samples were measured semi-quantitatively by dot-blots and quantitatively by enzyme-linked immunosorbant assays (ELISA).

RESULTS

Undifferentiated ADSC were negative for chondrogenic markers, as well as HLA-ABC and HLA-DR epitopes in immunofluorescence. In contrast, TGF-β3-induced pellet cultures showed both expression of chondrogenic differentiation markers, such as transcription factor 9 (Sox 9), collagen type IIa and aggrecan, and an up-regulation of HLA-DR, beginning at day 7 after induction. Interferon-γ (INF-γ) is known to up-regulate HLA-DR. Therefore we measured INF-γ levels in the supernatants of TGF-β3-induced pellets and, indeed, INF-γ was up-regulated during chondrogenesis in ADSC pellet cultures. However, both undifferentiated and TGF-β3-induced ADSC also showed expression of immunosuppressive HLA-G and interleukin (IL)-10 up-regulation.

CONCLUSIONS

These results suggest that the immunogenicity of adult stem cell-derived tissue should be tested in animal models before clinical trials for allogeneic engineered tissue are considered.

摘要

背景目的

间充质基质细胞（MSC）的软骨分化潜能及其免疫抑制特性已得到广泛研究。迄今为止，只有少数研究探讨了 MSC 在转分化后是否仍然保留其免疫抑制特性的问题。特别是，分化后免疫原性标志物（如人类白细胞抗原（HLA）-DR）的表达从未被研究过。

方法

使用 10ng/ml 转化生长因子（TGF）-β3 诱导来自 10 位不同患者的人脂肪组织来源的干细胞（ADSC）球培养物的软骨分化。在 28 天的时间过程中采集样本，并通过免疫组织化学和逆转录（RT）-聚合酶链反应（PCR）进行分析。通过斑点印迹和酶联免疫吸附试验（ELISA）定量分析样本上清液中的细胞因子水平。

结果

未分化的 ADSC 在免疫荧光中对软骨标志物以及 HLA-ABC 和 HLA-DR 表位呈阴性。相比之下，TGF-β3 诱导的球培养物在第 7 天诱导后开始表达软骨分化标志物，如转录因子 9（Sox9）、II 型胶原和聚集蛋白，以及 HLA-DR 的上调。干扰素-γ（INF-γ）已知上调 HLA-DR。因此，我们测量了 TGF-β3 诱导的球培养物上清液中的 INF-γ 水平，并且在 ADSC 球培养物的软骨形成过程中确实上调了 INF-γ。然而，未分化和 TGF-β3 诱导的 ADSC 也表现出免疫抑制 HLA-G 和白细胞介素（IL）-10 的上调。