Objective To propagate and titrate the adapted RT-SHIV from Chinese rhesus monkey using TZM-bl，CEMx174 and PBMCs cells，and to compare the possible difference of these viruses which propagated in PBMC and CEMx174，respectively. Methods Rhesus macaques free-living in China were infected with RT-SHIV intravenously.Blood samples were collected regularly and viral loads were monitored. Peripheral blood mononuclear cells ( PBMCs) were examined when the viral load reached the peak，and were co-cultured with PBMCs from uninfected rhesus or CEMx174 cells. The P24 antigen level from supernatant was monitored regularly. The co-cultured virus was collected，packed and frozen when the viral replication reaches the peak. The collected viruses as a stock were charactarized with the RNA viral loads，P24 antigen level and titration of TCID50. Results In this study，in tatol of 78 mL of RT-SHIV in monkeys’PBMCs and 85 mL of RT-SHIV was propagated in CEMx174 cells，respectively. The similarity between RT gene sequence and original sequence was 99% ，only two mutation of amino acid at 254 and 265. RT-SHIV ( PBMC) and RT-SHIV (CEMx174) viral loads were titrated with TZM-bl，CEMx174 and PBMCs，and their results were 1. 641 × 108 copies/mL and 8. 375 × 108 copies/mL. respectively，P24 antigenlevel were 20. 745 ng /mL and 4. 28 ng /mL，respectively，titrations of TCID50 in TZM-bl，CEMx174，PBMC were 3. 16 × 105TCID50 /mL and 1 × 104 TCID50 /mL;5 × 102 TCID50 /mL and 5 × 105 TCID50 /mL;5 × 102 TCID50 /mL and 5 × 103 TCID50 /mL，respectively. Conclusions RT-SHIV propagated from PBMCs has higher infective ablity than that from the other cells in vitro.