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Mesenchymal Stem Cell Induced In-Vitro Generation of Regulatory T-Cells – Future Novel Cellular Therapy for Tolerance Induction in Transplantation?

S. Dave, A. Vanikar, H. Trivedi

Department of Pathology, Laboratory Medicine, Transfusion Services and Immunohematology, Institute Of Kidney Diseases &
Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), Ahmedabad, Gujarat, India
Department of Nephrology and Transplantation Medicine, Institute Of Kidney Diseases &
Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), Ahmedabad, Gujarat, India

Meeting: 2013 American Transplant Congress

Abstract number: B1128

Introduction:

Regulatory T-cells (Tregs) have recently been recognized as key players in transplantation immunobiology with little knowledge about their source and dose for tolerance induction.

We present in-vitro model of induction of T-regs using donor adipose tissue derived mesenchymal stem cells (AD-MSC).

Material and Methodology:

Three grams subcutaneous adipose tissue was collected from 14 renal allograft donors during nephrectomy and cultured in lab using Α-minimal essential media. On 10 th day, AD-MSC were harvested. Peripheral blood mononuclear cells (PBMC) were derived from immunosuppressed recipients and were divided in two parts, one as responder-PBMC (R.PBMC) and other subjected to irradiation served as stimulator-PBMC (S.PBMC). R.PBMC and S.PBMC were layered on AD-MSC for 9 days in RPMI supplemented with IL-2. R.PBMC and S.PBMC co-cultured in absence of AD-MSC under similar conditions served as control. Cells were harvested and analysed for viability, suppression assay and phenotypic characterization using FACScan for CD127 -/low CD4+ CD25 high, CTLA-4+ and infused in thymus and portal circulation of recipients 3 weeks post-transplant.

Results:

Mean 4.4 ± 1.5 x102 AD-MSC/¯o;L were generated and co-cultured with mean 1.5 ± 1.6 x104 PBMC. Mean CD127-/low CD4+ CD25 high were increased from 3.11 ± 2.8 % to 25.1 ± 10% and CTLA-4+ increased from 24.2 ± 16.1 % to 48.3 ± 20.5 % at the end. Interestingly there was rise in CD4+ CD8+ cells from mean 5 ± 8.9 % to 48.9 ± 38.8 %. Tregs suppressive capacity was maintained in populations layered on AD-MSC for 12 days while control cells lost all suppressive activity after 3 days. Tregs infusion was uneventful. Over a mean follow-up of 49.07±27.28 days, all the patients are doing well with stable mean serum creatinine, 1.2 mg/dL without immune injury.

Conclusion:

We have successfully induced Tregs [CD127 -/low CD4+ CD25 high, CTLA-4+ and CD4+ CD8+ ] using donor AD-MSC from immunosuppressed recipient PBMC. This novel model will open up the gateway to tolerance in allo/autoimmunity.

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To cite this abstract in AMA style:

Dave S, Vanikar A, Trivedi H. Mesenchymal Stem Cell Induced In-Vitro Generation of Regulatory T-Cells – Future Novel Cellular Therapy for Tolerance Induction in Transplantation? [abstract]. Am J Transplant. 2013; 13 (suppl 5). https://atcmeetingabstracts.com/abstract/mesenchymal-stem-cell-induced-in-vitro-generation-of-regulatory-t-cells-future-novel-cellular-therapy-for-tolerance-induction-in-transplantation/. Accessed July 3, 2025.

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