Expansion of Sorted Cells with IL-2 and TAC

Expansion protocol was adapted from previous protocols with minor modifications.^{10 (link)} Sorted cells were expanded in Roswell Park Memorial Institute 1640 media supplemented with l-glutamine, penicillin/streptomycin, sodium pyruvate, and 10% of human AB pooled serum, in the presence of recombinant human IL-2 (1000 U/mL) (Novartis) and Dynabead Human T-activator CD3/CD28 (Life Technologies) in a 1:1 cell to bead ratio over two 7-day rounds of expansion. During the second round of expansion, 8 ng/mL TAC (Sigma) dissolved in dimethyl sulfoxide (DMSO) (Sigma), based on the published trough level in patients with stable liver or kidney allografts,^{6 (link),15 (link),16 (link)} or empty DMSO control added to cell media (final concentration of DMSO in culture was 1.92 × 10⁻⁴% volume).

Free full text: Click here

Arroyo Hornero R., Betts G.J., Sawitzki B., Vogt K., Harden P.N, & Wood K.J. (2016). CD45RA Distinguishes CD4+CD25+CD127−/low TSDR Demethylated Regulatory T Cell Subpopulations With Differential Stability and Susceptibility to Tacrolimus-Mediated Inhibition of Suppression. Transplantation, 101(2), 302-309.

Publication 2016

recombinant human IL-2 Dynabead Human T-activator CD3/CD28 dimethyl sulfoxide (DMSO)

Allografts Cell Dimethyl sulfoxide Glutamine Human Kidney Liver Patients Penicillin Pyruvate Recombinant human il 2 Serum Sodium Streptomycin

Corresponding Organization : University of Oxford

Top 5 similar protocols

Variable analysis

independent variables

Presence of TAC (8 ng/mL) dissolved in DMSO or empty DMSO control added to cell media during the second round of expansion

dependent variables

Expansion of sorted cells

control variables

Roswell Park Memorial Institute 1640 media supplemented with L-glutamine, penicillin/streptomycin, sodium pyruvate, and 10% of human AB pooled serum
Presence of recombinant human IL-2 (1000 U/mL)
Presence of Dynabead Human T-activator CD3/CD28 in a 1:1 cell to bead ratio
Two 7-day rounds of expansion

negative control

Empty DMSO control added to cell media

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!