Preliminary Study of Single Cell Isolation and Single Cell PCR in Lymph Node Germinal Centers / 复旦学报（医学版）

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.