Clinical-grade Treg cell manufacturing

1. Clinical-grade Treg separation
 

With Treg cells being more and more used as treatment options to suppress solid organ rejection after transplantation, for autoimmune diseases, and as CAR Treg cells, the demand for ultra-pure antigen-specific Tregs has recently increased tremendously. 

Miltenyi Biotec now offers solutions to meet the needs of the field. The combination of the CliniMACS Prodigy for pre-enrichment and the MACSQuant Tyto Cell Sorter for subsequent high-purity cell sorting guarantees full sterility of your medical product.  

2. Clinical-grade Treg cell expansion
 

In order to improve the purity and quantity of clinical-grade Treg cells, we offer GMP-compatible expansion products. Ideal cultivation and expansion of Treg cells rely on the strong synergy of:

• MACS GMP ExpAct Treg Kit
• TexMACS™ GMP Medium,
• MACS GMP Cytokines and
• MACS GMP Rapamycin

MACS GMP ExpAct Treg Kit

The MACS GMP ExpAct Treg Kit has been designed to efficiently expand Treg cells after a CliniMACS CD25 enrichment procedure while maintaining FoxP3 expression. It can be used in combination with TexMACS GMP Medium and MACS GMP Rapamycin, which inhibits the expansion of conventional T cells and thereby enhances the proportion of expanded Treg cells in culture.

MACS GMP Rapamycin

Profit from MACS GMP Rapamycin to enhance the proportion of FoxP3⁺ Treg cells and to inhibit the expansion of conventional T cells during the cultivation and expansion step. We offer 100 nM ready-to-use MACS GMP Rapamycin to improve your culturing.
 

TexMACS GMP Medium

To enhance your Treg cell culturing, we offer TexMACS GMP medium which is specifically developed for T cell cultivation. TexMACS GMP medium is serum- and xeno-component free, but it is supplemented with pharmaceutical-grade human serum albumin. To support you with the consistently high standards, a QC functionality test is done on every batch.
 

MACS GMP Cytokines (IL-2)

Our MACS GMP Cytokines are designed according to the recommendations of <USP 1043> on ancillary materials and manufactured and tested under a certified ISO 13485 quality system. In addition, we provide you with a lot-to-lot consistency and lot-specific certificates of analysis. 
 

3. In-process and quality control 
 

The Treg Suppression Inspector has been developed for the in vitro functional characterization of human CD4⁺ CD25⁺ regulatory T cells.
 

Treg Detection Kits  

Our Treg Detection Kits are optimized antibody kits with pre-titrated antibodies that ensure a reliable flow cytometric analysis of Treg cells. The kits are ready-to-use cocktails, which conveniently reduce pipetting steps. 
 

REAfinity™ Recombinant Antibodies 

REAfinity Antibodies are recombinant antibodies that provide superior lot-to-lot consistency and purity compared to mouse or rat hybridoma-derived, monoclonal antibodies. They have been recombinantly engineered to produce highly specific antibodies that require no FcR blocking step. Additionally, they all have the same IgG1 isotype, requiring less isotype controls.
 

References

1. Martelli, M. F. et al. (2014) HLA‑haploidentical transplantation with regulatory and conventional T‑cell adoptive immunotherapy prevents acute leukemia relapse. Blood 124(4): 638–644
2. Martelli, M. F. et al. (2015) Next generation HLA-haploidentical HSCT. Bone Marrow Transplant. 50: 63–66.
   
3. Theil, A. et al. (2015) Adoptive transfer of allogeneic regulatory T cells into patients with chronic graft‑versus‑host disease. Cytotherapy 17(4): 473–486.
   
4. Hannon, M. et al. (2014) Infusion of clinical‑grade enriched regulatory T cells delays experimental xenogeneic graft‑versus‑host disease. Transfusion 54(2): 353–363.
   
5. Nadig, S.N. et al. (2010) In vivo prevention of transplant arteriosclerosis by ex vivo-expanded human regulatory T cells. Nat. Med. 16: 809.
   
6. Sagoo, P. et al. (2011) Human regulatory T cells with alloantigen specificity are more potent inhibitors of alloimmune skin graft damage than polyclonal regulatory T cells. Sci. Transl. Med. 3: 83ra42.
   
7. Issa, F. et al. (2010) Ex vivo expanded human regulatory T cells prevent the rejection of skin allografts in a humanized mouse model. Transplantation 90: 1321.
   
8. Wu, D.C. et al. (2013) Ex vivo expanded human regulatory T cells can prolong survival of a human islet allograft in a humanized mouse model. Transplantation 96: 707.
   
9. Safina, N. et al. (2016) Successful expansion of functional and stable regulatory T cells for immunotherapy in liver transplantation. Oncotarget 7(7): 7563–7577.
   
10. Bluestone J.A. (2015) Type 1 diabetes immunotherapy using polyclonal regulatory T cells. Sci. Transl. Med. 7, 315ra189.
    
11. Putnam A. L. et al. (2013) Clinical grade manufacturing of human alloantigen-reactive regulatory T cells for use in transplantation. A. J. Transplant. 13: 3010–3020.
    
12. Battaglia, M. et al. (2005) Rapamycin selectively expands CD4+CD25+FoxP3+ regulatory T cells. Blood 105: 4743–4748.