Document Type : Original research articles
Authors
1
Stem Cell Research Group, Medical Research Centre of Excellence, National Research Centre, Cairo, Egypt.
2
Department of Medical Molecular Genetics, Human Genetics and Genome Research Division, National Research Centre, Cairo, Egypt
3
Stem Cell Research Group, Medical Research Centre of Excellence, National Research Centre, Cairo, Egypt
4
Department of Molecular Genetics and Enzymology, Human Genetics and Genome Research Division, National Research Centre, Cairo, Egypt
5
Department of Immunogenetics, Human Genetics and Genome Research Division, National Research Centre, Cairo, Egypt
6
Department of Endocrine Surgery, National Institute of Diabetes and Endocrinology, Cairo, Egypt
7
Department of Reproductive Health Research, Medical Research Division, National Research Centre, Cairo, Egypt
8
Egypt Center for Research and Regenerative Medicine, Cairo, Egypt
9
Department of Biochemistry and Molecular Biology, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt
Abstract
Adipose tissue is a readily available and plentiful source of multipotent mesenchymal stromal/stem cells (AT-MSC). The immunomodulatory properties of AT-MSC are being introduced in type 2 diabetes (T2D) cell- based therapy. The study aimed to uncover the impact of T2D, on the interplay between AT-MSC and immune cells to develop an effective and safe AT-MSC immunotherapeutic modality. Thus, a direct allogenic co-culture of healthy AT-MSC (nAT-MSC) and peripheral blood mononuclear cells (PBMC), from healthy (nPBMC) or T2D (dPBMC) donors, and stimulated with anti-CD3/CD28, was established in vitro. PBMC proliferation was evaluated by measuring 5-bromo-20-deoxyuridine (Brdu) incorporation in the DNA of proliferating cells in a colorimetric ELISA assay. CD3+ T cell activation surface markers (CD25 and HLA-DR) expression was detected using a flow cytometer. As well, the anti-proliferative effect of naïve and interferon gamma (IFN-ɤ) - primed AT-MSC, isolated from T2D patients (dAT-MSC), on autologous PBMC was explored using the Brdu proliferation assay. In the applied co-culture setting, we observed that the diabetic milieu did not significantly impact the potential of nAT-MSC to suppress stimulated PBMC proliferation. However, it significantly compromised nAT-MSC ability to modulate the activation markers expression, making them less potent to suppress CD25 and HLA-DR expression. Moreover, the dAT-MSC had attenuated ability to suppress the proliferation of autologous stimulated dPBMC, nevertheless, priming of dAT-MSC with IFN-ɤ, could, to an extent, improve such defect. The results suggest that T2D might affect the immunosuppressive potential of AT-MSC and pre-conditioning of dAT-MSC with a pro-inflammatory stimuli could enhance their therapeutic effect.
Keywords
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