Exploration on factors influencing HLA-C molecular expression level by flow cytometry
[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34+ cell counting detection. The influen...
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Institute of Blood Transfusion of Chinese Academy of Medical Sciences
2025-01-01
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| Series: | Zhongguo shuxue zazhi |
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| Online Access: | https://www.cjbt.cn/thesisDetails#10.13303/j.cjbt.issn.1004-549x.2025.01.013&lang=en |
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| _version_ | 1832585131294982144 |
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| author | LI Yunan JIANG Renhui CAI Siqi LIU Jie DENG Zhihui |
| author_facet | LI Yunan JIANG Renhui CAI Siqi LIU Jie DENG Zhihui |
| author_sort | LI Yunan |
| collection | DOAJ |
| description | [Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34+ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results. |
| format | Article |
| id | doaj-art-6996022e44d74507ab71e78503398da2 |
| institution | Kabale University |
| issn | 1004-549X |
| language | zho |
| publishDate | 2025-01-01 |
| publisher | Institute of Blood Transfusion of Chinese Academy of Medical Sciences |
| record_format | Article |
| series | Zhongguo shuxue zazhi |
| spelling | doaj-art-6996022e44d74507ab71e78503398da22025-01-27T01:46:11ZzhoInstitute of Blood Transfusion of Chinese Academy of Medical SciencesZhongguo shuxue zazhi1004-549X2025-01-01381798410.13303/j.cjbt.issn.1004-549x.2025.01.0131004-549X(2025)1-0079-06Exploration on factors influencing HLA-C molecular expression level by flow cytometryLI Yunan0JIANG Renhui1CAI Siqi2LIU Jie3DENG Zhihui4Institute of Blood Transfusion, Shenzhen Blood Center, Shenzhen 518040, ChinaInstitute of Blood Transfusion, Shenzhen Blood Center, Shenzhen 518040, ChinaDepartment of Transfusion, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518033, ChinaInstitute of Blood Transfusion, Shenzhen Blood Center, Shenzhen 518040, ChinaInstitute of Blood Transfusion, Shenzhen Blood Center, Shenzhen 518040, China[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34+ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.https://www.cjbt.cn/thesisDetails#10.13303/j.cjbt.issn.1004-549x.2025.01.013&lang=enhuman leukocyte antigen (hla)hla-c geneexpression levelflow cytometrymean fluorescence intensity (mfi) |
| spellingShingle | LI Yunan JIANG Renhui CAI Siqi LIU Jie DENG Zhihui Exploration on factors influencing HLA-C molecular expression level by flow cytometry Zhongguo shuxue zazhi human leukocyte antigen (hla) hla-c gene expression level flow cytometry mean fluorescence intensity (mfi) |
| title | Exploration on factors influencing HLA-C molecular expression level by flow cytometry |
| title_full | Exploration on factors influencing HLA-C molecular expression level by flow cytometry |
| title_fullStr | Exploration on factors influencing HLA-C molecular expression level by flow cytometry |
| title_full_unstemmed | Exploration on factors influencing HLA-C molecular expression level by flow cytometry |
| title_short | Exploration on factors influencing HLA-C molecular expression level by flow cytometry |
| title_sort | exploration on factors influencing hla c molecular expression level by flow cytometry |
| topic | human leukocyte antigen (hla) hla-c gene expression level flow cytometry mean fluorescence intensity (mfi) |
| url | https://www.cjbt.cn/thesisDetails#10.13303/j.cjbt.issn.1004-549x.2025.01.013&lang=en |
| work_keys_str_mv | AT liyunan explorationonfactorsinfluencinghlacmolecularexpressionlevelbyflowcytometry AT jiangrenhui explorationonfactorsinfluencinghlacmolecularexpressionlevelbyflowcytometry AT caisiqi explorationonfactorsinfluencinghlacmolecularexpressionlevelbyflowcytometry AT liujie explorationonfactorsinfluencinghlacmolecularexpressionlevelbyflowcytometry AT dengzhihui explorationonfactorsinfluencinghlacmolecularexpressionlevelbyflowcytometry |