Impact of culture conditions on the proliferative lifespan of human T cells in vitro

Röth, Alexander; Schneider, Lars; Himmelreich, Heike; Baerlocher, Gabriela M.

doi:10.1080/14653240601113197

Cited by 11 publications

(9 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…about 1000-fold expansion, which is consistent with previous observations using this culture method (31,32). Following 12 population doublings, an increase in IL-2 stimulation or in the number of beads did not allow for further growth (supplemental Fig.…”

Section: Global Characteristics Of Age-associated Protein Expression supporting

confidence: 80%

Predicting Cytotoxic T-cell Age from Multivariate Analysis of Static and Dynamic Biomarkers

Rivet

Hill

et al. 2011

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Adoptive T-cell transfer therapy relies upon in vitro expansion of autologous cytotoxic T cells that are capable of tumor recognition. The success of this cell-based therapy depends on the specificity and responsiveness of the T cell clones before transfer. During ex vivo expansion, CD8؉ T cells present signs of replicative senescence and loss of function. The transfer of nonresponsive senescent T cells is a major bottleneck for the success of adoptive T-cell transfer therapy. Quantitative methods for assessing cellular age and responsiveness will facilitate the development of appropriate cell expansion and selection protocols. Although several biomarkers of lymphocyte senescence have been identified, these proteins in isolation are not sufficient to determine the age-dependent responsiveness of T cells. We have developed a multivariate model capable of extracting combinations of markers that are the most informative to predict cellular age. To acquire signaling information with high temporal resolution, we designed a microfluidic chip enabling parallel lysis and fixation of stimulated cell samples on-chip. The acquisition of 25 static biomarkers and 48 dynamic signaling measurements at different days in culture, integrating single-cell and population based information, allowed the multivariate regression model to accurately predict CD8؉ T-cell age. From surface marker expression and early phosphorylation events following T-cell receptor stimulation, the model successfully predicts days in culture and number of population doublings with R 2 ‫؍‬ 0.91 and 0.98, respectively. Furthermore, we found that impairment of early signaling events following T cell receptor stimulation because of long term culture allows prediction of costimulatory molecules CD28 and CD27 expression levels and the number of population divisions in culture from a limited subset of signaling proteins. The multivariate analysis highlights the information content of both averaged biomarker values and heterogeneity metrics for prediction of cellular age within a T cell population.

show abstract

Section: Global Characteristics Of Age-associated Protein Expression supporting

confidence: 80%

Predicting Cytotoxic T-cell Age from Multivariate Analysis of Static and Dynamic Biomarkers

Rivet

Hill

et al. 2011

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…For adoptive transferring of T lymphocyte, a large number of cells are required, and on the other hand, effi ciently, adoptive transfer need an antigenspecifi c T cell obtaining from blood or tissue sites, but the obtained cells are not suffi cient [4]. The abovementioned methods were usually used to effi ciently expand these cells for the next investigation [11]. The aim of this study is to defi ne proliferation ability of two different stimulators on CD4+ lymphocytes.…”

mentioning

confidence: 99%

Extensive Proliferation of Cd4+ Lymphocyte by Both Phytohaemagglutinin A and Anti-Cd2/Cd3/ Cd28 Macsibeads

Mirzakhani

Shahbazi

Darvish

et al. 2018

Acta Medica Bulgarica

View full text Add to dashboard Cite

Background: Lymphocytes proliferate considerably following appropriate stimulation in vitro. Autologous T cells are obtained from whole blood or tissue sites in relatively limited amounts. We need a method to expand these cells efficiently, study their functions and manipulate them to create appropriate cells for transferring to the patient with infection and cancer. Objectives: The aim of this study is to determine proliferation ability of two different stimulators on CD4+ lymphocytes. Methods: Lymphocytes were isolated from blood samples of healthy donors after removing adherent cells (monocytes).The efficacy of MACSiBead™ coated with anti-CD2, anti-CD3, anti-CD28 (anti-CD2/CD3/CD28) was compared with Phytohaemagglutinin A (PHA) on CD4+ lymphocytes proliferation using carboxyfluorescein diacetate succinimidyl ester (CFSE) in cell culture media. The percentage of proliferating cells was analyzed using flow cytometry. Results: Both stimulators induced extensive proliferation of CD4+ lymphocytes but proliferation ability of PHA was higher compared to stimulation by anti-CD2/CD3/CD28 MACSiBead™. The proliferation rate of cells stimulated by PHA was 93.8% ± 3.37% whereas it was 85.2% ± 4.7% in cells stimulated by anti-CD2/CD3/CD28 MACSiBead™. Conclusions: Our results show that MACSiBead™ along with PHA can be used to obtain a large number of expanded CD4+ lymphocytes.

show abstract

“…As biologic alternatives, AS and AP have belong the first proposed options to supplement the cell media. It is reported that AS outperforms FBS for the cultivation of human T cells [20] as well as human articular chondrocytes [21]. AS and AP have been shown to be suitable supplements also for the expansion of various stem cells obtained from different origins without adversely affecting their differentiation capacity [22][23][24][25].…”

Section: Discussionmentioning

confidence: 99%

Autologous plasma versus fetal calf serum as a supplement for the culture of neutrophils.

Alipour

Fatemi

Alsahebfosul

et al. 2019

Preprint

View full text Add to dashboard Cite

Objective Currently the replacement of fetal calf serum (FCS) by a more suitable alternative is a sought aim in the field of tissue and cell culture research. Autologous plasma (AP) and especially autologous serum (AS) have been shown to be effective substitutes of FCS in culture media for some of cell types. Nevertheless, there is no comparative data on the most appropriate supplement for cell media in neutrophil studies, it is now unclear whether AP have relatively an equal, superior or inferior performance to FCS in neutrophil cell culture. In the present study, human blood neutrophils were isolated and cultured in FCS- or AP-supplemented medium. After 12, 36 and 60 hours of incubation, cell viability, oxidative burst and CD11b expression were determined by flow cytometry.Results Compared to the culture of neutrophils in FCS 10% medium, the culture of neutrophils in a medium with AP 10% could prolong their life span without affecting their function. The findings introduce AP as a better supplement for human neutrophil cell culture than FCS and propose a simple and economical procedure for neutrophil isolation and culture.

show abstract

Impact of culture conditions on the proliferative lifespan of human T cells in vitro

Cited by 11 publications

References 44 publications

Predicting Cytotoxic T-cell Age from Multivariate Analysis of Static and Dynamic Biomarkers

Predicting Cytotoxic T-cell Age from Multivariate Analysis of Static and Dynamic Biomarkers

Extensive Proliferation of Cd4+ Lymphocyte by Both Phytohaemagglutinin A and Anti-Cd2/Cd3/ Cd28 Macsibeads

Autologous plasma versus fetal calf serum as a supplement for the culture of neutrophils.

Contact Info

Product

Resources

About