Preliminary Study of Single Cell Isolation and Single Cell PCR in Lymph Node Germinal Centers
- VernacularTitle:单细胞分离及单细胞PCR技术在淋巴结生发中心研究中的应用
- Author:
Wentao YANG
;
Liangzhong XU
- Publication Type:Journal Article
- Keywords:
single cell isolation;
polymerase chain reaction;
lymph node
- From:
Fudan University Journal of Medical Sciences
2000;27(6):460-463
- CountryChina
- Language:Chinese
-
Abstract:
Purpose To explore the value of single cell isolation from tissue slides and single cell polymerase chain reaction(PCR) in the study of lymphy node germinal centers. Methods By single-cell isolation and single-cell PCR,rearranged immunoglobulin(Ig) genes were amplified from single Ki67(+) cells and single CD3(+) cells.The Ⅴ-region family specific primers were designed to Ig heavy chain leader region and light chain κ and λ framework region Ⅰ. Results PCR efficiency of Ki67(+) cells were 18.7% and 50.0% respectively in 2 lymph nodes.However,in 40 CD3(+) T cells,no Ig gene rearrangements were observed,which confirmed the efficiency and specificity of single cell isolation and single cell PCR. Conclusions Our study demonstrated the feasibility of single cell isolation from tissue slides and single cell PCR.This makes a sound basis for the use of technique in the further study of lymphoid diseases.
