Flow cytometric analyses of the viability, surface marker expression and function of lymphocytes from children following cryopreservation

Chen,Xi; Zhang,Hui; Mou,Wenjun; Qi,Zhan; Ren,Xiaoya; Wang,Guoliang; Jiao,Hong; Kong,Xiaohui; Gui,Jingang

doi:10.3892/mmr.2016.5780

Flow cytometric analyses of the viability, surface marker expression and function of lymphocytes from children following cryopreservation

Authors:
- Xi Chen
- Hui Zhang
- Wenjun Mou
- Zhan Qi
- Xiaoya Ren
- Guoliang Wang
- Hong Jiao
- Xiaohui Kong
- Jingang Gui
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Affiliations: Key Laboratory of Major Diseases in Children by Ministry of Education, Beijing Children's Hospital affiliated with Capital Medical University, Beijing 100045, P.R. China
Published online on: September 26, 2016 https://doi.org/10.3892/mmr.2016.5780
Pages: 4301-4308

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Abstract

Flow cytometric analysis is important for the investigation and clinical preparation of lymphocytes from children. However, the strict requirement of cell freshness and inter‑assay variability limits the application of this methodology for pediatric investigations. Therefore, it is necessary to identify a reliable cryopreservative method capable of maintaining high cell viability and proper cell function in lymphocytes from children. In the present study, eight commonly‑used cell cyropreservative methods were used, and their effects on cell viability, surface marker expression and cell function were examined. In addition, how these methods affect the distribution of T‑cell receptor Vβ subfamilies were also determined. The results of the present study provided valuable experimental evidence, based on which the optimal method for the cryopreservation of lymphocytes from children in pediatric investigations and clinical applications can be selected.

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1	Chattopadhyay PK and Roederer M: Cytometry: Today's technology and tomorrow's horizons. Methods. 57:251–258. 2012. View Article : Google Scholar : PubMed/NCBI
2	Buhl T, Legler TJ, Rosenberger A, Schardt A, Schön MP and Haenssle HA: Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy. Cancer Immunol Immunother. 61:2021–2031. 2012. View Article : Google Scholar : PubMed/NCBI
3	Finn OJ: Cancer Immunology. N Engl J Med. 358:2704–2715. 2008. View Article : Google Scholar : PubMed/NCBI
4	Curran KJ, Pegram HJ and Brentjens RJ: Chimeric antigen receptors for T cell immunotherapy: Current understanding and future directions. J Gene Med. 14:405–415. 2012. View Article : Google Scholar : PubMed/NCBI
5	Antoniewicz-Papis J, Lachert E, Woźniak J, Janik K and Łętowska M: Methods of freezing cord blood hematopoietic stem cells. Transfusion. 54:194–202. 2014. View Article : Google Scholar : PubMed/NCBI
6	Stevens VL, Patel AV, Feigelson HS, Rodriguez C, Thun MJ and Calle EE: Cryopreservation of whole blood samples collected in the field for a large epidemiologic study. Cancer Epidemiol Biomarkers Prev. 16:2160–2163. 2007. View Article : Google Scholar : PubMed/NCBI
7	Germann A, Schulz JC, Kemp-Kamke B, Zimmermann H and von Briesen H: Standardized serum-free cryomedia maintain peripheral blood mononuclear cell viability, recovery, and antigen-specific T-cell response compared to fetal calf serum-based medium. Biopreserv Biobank. 9:229–236. 2011. View Article : Google Scholar : PubMed/NCBI
8	Dijkstra-Tiekstra MJ, Setroikromo AC, Kraan M, Gkoumassi E and de Wildt-Eggen J: Optimization of the freezing process for hematopoietic progenitor cells: Effect of precooling, initial dimethyl sulfoxide concentration, freezing program, and storage in vapor-phase or liquid nitrogen on in vitro white blood cell quality. Transfusion. 54:3155–3163. 2014. View Article : Google Scholar : PubMed/NCBI
9	Petrenko YA, Rogulska OY, Mutsenko VV and Petrenko AY: A sugar pretreatment as a new approach to the Me2SO- and xeno-free cryopreservation of human mesenchymal stromal cells. Cryo Letters. 35:239–246. 2014.PubMed/NCBI
10	Hreinsson J, Zhang P, Swahn ML, Hultenby K and Hovatta O: Cryopreservation of follicles in human ovarian cortical tissue. Comparison of serum and human serum albumin in the cryoprotectant solutions. Hum Reprod. 18:2420–2428. 2003. View Article : Google Scholar : PubMed/NCBI
11	Wiegering V, Eyrich M, Wunder C, Günther H, Schlegel PG and Winkler B: Age-relate changes in intracellular cytokine expression in healthy children. Eur Cytokine Netw. 20:75–80. 2009.PubMed/NCBI
12	Bunders M, Cortina-Borja M and Newell ML: European Collaborative Study: Age-related standards for total lymphocyte, CD4+ and CD8+ T cell counts in children born in Europe. Pediatr Infect Dis J. 24:595–600. 2005. View Article : Google Scholar : PubMed/NCBI
13	Wu J, Liu D, Tu W, Song W and Zhao X: T-cell receptor diversity is selectively skewed in T-cell populations of patients with Wiskott-Aldrich syndrome. J Allergy Clin Immunol. 135:209–216. 2015. View Article : Google Scholar : PubMed/NCBI
14	Tzifi F, Kanariou M, Tzanoudaki M, Mihas C, Paschali E, Chrousos G and Kanaka-Gantenbein C: Flow cytometric analysis of the CD4+ TCR Vβ repertoire in the peripheral blood of children with type 1 diabetes mellitus, systemic lupus erythematosus and age-matched healthy controls. BMC Immunol. 14:332013. View Article : Google Scholar : PubMed/NCBI
15	Attaf M, Huseby E and Sewell AK: αβ T cell receptors as predictors of health and disease. Cell Mol Immunol. 12:391–399. 2015. View Article : Google Scholar : PubMed/NCBI
16	Gil A, Yassai M, Naumov Y and Selin L: Narrowing of human influenza A virus-specific T cell receptor α and β repertoires with increasing age. J Virol. 89:4102–4116. 2015. View Article : Google Scholar : PubMed/NCBI
17	Osugi Y, Hara J, Kurahashi H, Sakata N, Inoue M, Yumura-Yagi K, Kawa-Ha K, Okada S and Tawa A: Age-related changes in surface antigens on peripheral lymphocytes of healthy children. Clin Exp Immunol. 100:543–548. 1995. View Article : Google Scholar : PubMed/NCBI
18	Chen H, Ndhlovu ZM, Liu D, Porter LC, Fang JW, Darko S, Brockman MA, Miura T, Brumme ZL, Schneidewind A, et al: TCR clonotypes modulate the protective effect of HLA class I molecules in HIV-1infection. Nat Immunol. 13:691–700. 2012. View Article : Google Scholar : PubMed/NCBI
19	Aleman K, Noordzij JG, de Groot R, van Drogen JJ and Hartwig NG: Reviewing Omenn syndrome. Eur J Pediatr. 160:718–725. 2001. View Article : Google Scholar : PubMed/NCBI
20	Somma P, Ristori G, Battistini L, Cannoni S, Borsellino G, Diamantini A, Salvetti M, Sorrentino R and Fiorillo MT: Characterization of CD81 T cell repertoire in identical twins discordant and concordant for multiple sclerosis. J Leukoc Biol. 81:696–710. 2007. View Article : Google Scholar : PubMed/NCBI
21	Fang H, Yamaguchi R, Liu X, Daigo Y, Yew PY, Tanikawa C, Matsuda K, Imoto S, Miyano S and Nakamura Y: Quantitative T cell repertoire analysis by deep cDNA sequencing of T cell receptor α and β chains using next-generation sequencing (NGS). Oncoimmunology. 3:e9684672015. View Article : Google Scholar : PubMed/NCBI
22	Gao F and Wang K: Ligation-anchored PCR unveils immune repertoire of TCR-beta from whole blood. BMC Biotechnol. 15:392015. View Article : Google Scholar : PubMed/NCBI