Comparative Study of the Two High-Efficient Strategies for in vitro Generation of Human Umbilical Cord Blood-derived Natural Killer Cells.
10.19746/j.cnki.issn.1009-2137.2023.02.035
- Author:
Ti-Er WANG
1
;
Yun-Yan SUN
2
;
Zhong-Chao HAN
3
;
Lei-Sheng ZHANG
4
;
Ming-Xia SHI
5
Author Information
1. Department of Hematology, The First Affiliated Hospital of Kunming Medical University, Hematology Research Center of Yunnan Province, Kunming 650032,Yunnan Province, China.
2. Department of Hematology, The First Affiliated Hospital of Kunming Medical University, Hematology Research Center of Yunnan Province, Kunming 650032,Yunnan Province, China,Department of Hematology, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University,Yunnan Cancer Center, Kunming 650118, Yunnan Province, China.
3. 3Key Laboratory of Gastrointestinal Tumor Diagnosis and Therapy of National Health Commission & Key Laboratory of Molecular Diagnostics and Precision Medicine for Surgical Oncology in Gansu Province, Gansu Provincial Hospital, Lanzhou 730000, Gansu Province, China.
4. Key Laboratory of Gastrointestinal Tumor Diagnosis and Therapy of National Health Commission & Key Laboratory of Molecular Diagnostics and Precision Medicine for Surgical Oncology in Gansu Province, Gansu Provincial Hospital, Lanzhou 730000, Gansu Province, China,Key Laboratory of Radiation Technology and Biophysics, Hefei Institute of Physical Science, Chinese Academy of Sciences, Hefei 230031, Anhui Province, China,E-mail：leisheng_zhang@163.com.
5. Department of Hematology, The First Affiliated Hospital of Kunming Medical University, Hematology Research Center of Yunnan Province, Kunming 650032,Yunnan Province, China,E-mail: shmxia2002@sina.com.
- Publication Type:Journal Article
- Keywords:
NK cells;
activation and amplification;
cell subpopulation;
cell vitality;
cytotoxicity
- MeSH:
Humans;
Fetal Blood;
Killer Cells, Natural;
T-Lymphocytes;
Leukocytes, Mononuclear/metabolism*;
Cell Proliferation;
CD56 Antigen/metabolism*
- From:
Journal of Experimental Hematology
2023;31(2):553-561
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the similarities and variations of biological phenotype and cytotoxicity of human umbilical cord blood natural killer cells (hUC- NK) after human umbilical cord blood-derived mononuclear cells (hUC-MNC) activated and expanded by two in vitro high-efficient strategies.
METHODS:Umbilical cord blood mononuclear cells (MNC) from healthy donor were enriched by Ficoll-based density gradient centrifugation. Then, the phenotype, subpopulations, cell viability and cytotoxicity of NK cells derived from Miltenyi medium (denoted as M-NK) and X-VIVO 15 (denoted as X-NK) were compared using a "3IL" strategy.
RESULTS:After a 14-day's culture, the contents of CD3-CD56+ NK cells were elevated from 4.25%±0.04% (d 0) to 71%±0.18% (M-NK) and 75.2%±1.1% (X-NK) respectively. Compared with X-NK group, the proportion of CD3+CD4+ T cells and CD3+CD56+ NKT cells in M-NK group decreased significantly. The percentages of CD16+, NKG2D+, NKp44+, CD25+ NK cells in X-NK group was higher than those in the M-NK group, while the total number of expanded NK cells in X-NK group was half of that in M-NK group. There were no significant differences between X-NK and M-NK groups in cell proliferation and cell cycle, except for the lower percentage of Annexin V+ apoptotic cells in M-NK group. Compared with X-NK group, the proportion of CD107a+ NK cells in M-NK group were higher under the same effector-target ratio (E∶T) (P<0.05).
CONCLUSION:The two strategies were adequate for high-efficient generation of NK cells with high level of activation in vitro, however, there are differences in biological phenotypes and tumor cytotoxicity.
