uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
Purification of regulatory T cells with the use of a fully enclosed high-speed microfluidic system
Show others and affiliations
2014 (English)In: Cytotherapy, ISSN 1465-3249, E-ISSN 1477-2566, Vol. 16, no 10, 1384-1389 p.Article in journal (Refereed) Published
Abstract [en]

Background aims. Despite promising advances in cellular therapies, it will be difficult to fully test or implement new therapies until advances are made in the processes for cell preparation. This study describes the use of an advanced prototype of a flow-cytometry cell purification system constructed for operation in a clinical environment to prepare regulatory T cells defined as CD4(+)/CD25(bright)/CD127(neg/low). Methods. The sort performance of the Gigasort system was directly compared with available droplet sorters using mixtures of highly fluorescent and non-fluorescent 5-mu m polystyrene particles. CD4(+)-enriched cell preparations were processed with the use of a sterile, disposable fluid handling unit with a chip containing parallel microfluidic-based sorters. Results. Similar purity and sort efficiency as found with droplet sorters were obtained with the 24-channel chip sorter system. Starting with 450 million fresh peripheral blood mononuclear cells, 150,000 to 1.7 million cells that were, on average, 85% FoxP3-positive and 97% viable, were obtained in <4 h. Conclusions. This study presents a technology adapted to regulatory requirements for clinical cell purification and that achieves high throughput and cell-friendly conditions by use of a microfluidic chip with 24 parallel microsorters, providing a rapid, sterile method of purifying regulatory T cells accurately and with excellent viability.

Place, publisher, year, edition, pages
2014. Vol. 16, no 10, 1384-1389 p.
National Category
Immunology in the medical area
URN: urn:nbn:se:uu:diva-220872DOI: 10.1016/j.jcyt.2014.05.016ISI: 000342396800007OAI: oai:DiVA.org:uu-220872DiVA: diva2:706829
Available from: 2014-03-21 Created: 2014-03-21 Last updated: 2014-11-05Bibliographically approved
In thesis
1. Preparatory Studies to Introduce Regulatory T Cells in Clinical Transplantation
Open this publication in new window or tab >>Preparatory Studies to Introduce Regulatory T Cells in Clinical Transplantation
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Solid organ transplantation has evolved from being an experimental procedure to a life-saving treatment for patients with end-stage organ failure. The risk of losing a transplant due to acute rejection is very low with the use of modern immunosuppressive protocols and the short-term results are impressive. However, long-term outcomes are suboptimal and transplant recipients are at increased risks for severe complications such as cancers, opportunistic infections and cardiovascular events. The previous struggle to achieve short-term survival has turned into a search for new strategies to improve patient and transplant longevity.

Regulatory T cells (TRegs), a subset of T cells, occur naturally in the immune system and have the capacity to down regulate immune responses. Under normal conditions they maintain self-tolerance and prevent excessive immune activation. Functional TReg defects lead to a massive autoimmune response and are not compatible with life. Preclinical data support that TRegs can be used as a cell therapy to prevent transplant rejection, with the potential to minimize the need for traditional immunosuppression and improve the long-term outcome.

This thesis aims to enhance the translation of TReg cell therapy to clinical organ transplantation. In particular, strategies for isolation and expansion of TRegs from uremic patients awaiting kidney transplantation have been assessed. A non-invasive imaging technique to study T cell products after intravenous administration was developed, for use in future clinical trials. The performance of a novel cell purification technique was investigated to potentially improve the clinical production of TRegs.

The thesis demonstrates that TRegs can be isolated and expanded from uremic patients to display potent suppressive properties in vitro. The mode of isolation and expansion affect the functional characteristics, where cells purified with cytometry based techniques and expanded with mature dendritic cells were the most advantageous. T cells can be labeled using the radioactive tracer [111In]oxine with preserved viability and subsequently followed in vivo with SPECT/CT for more than 1 week after intravenous administration. The use of microfluidic switch technology offers a novel way of purifying TRegs at high speed, purity and viability, under conditions compatible with clinical use.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. 80 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 983
Transplantation, Tolerance, Cell therapy, Regulatory T cell, In vivo imaging, Cell purification
National Category
Immunology in the medical area
urn:nbn:se:uu:diva-220873 (URN)978-91-554-8907-6 (ISBN)
Public defence
2014-05-10, Sal X, Universitetshuset, Biskopsgatan 3, Uppsala, 09:15 (English)
Available from: 2014-04-15 Created: 2014-03-21 Last updated: 2014-04-29

Open Access in DiVA

No full text

Other links

Publisher's full text

Search in DiVA

By author/editor
Korsgren, OlleBerglund, David
By organisation
Clinical ImmunologyTransplantation Surgery
In the same journal
Immunology in the medical area

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 503 hits
ReferencesLink to record
Permanent link

Direct link