Abstract: Objective To establish the isolation and culture system of human perinatal mesenchymal stem cells(P-MSCs), and explore the work flow of P-MSCs cell bank. Methods Our department started to store perinatal mesenchymal stem cells(P-MSCs) from October 2017. The umbilical cord and placenta tissues were collected after delivery in our hospital, then the umbilical cord mesenchymal stem cells, amniotic mesenchymal stem cells, and maternal mesenchymal stem cells were separated. After completion of safety and quality control testing on P-MSCs, we stored them in our cell bank. At present, there were various types of stem cells from 109 donors had been stored. Results Various types of P-MSCs were obtained, and all these P-MSCs had spindle shape and highly expressed stem cell positive markers, such as CD44, CD73 and CD90, while the expression of HLA-DR and CD34 proteins was relatively low. Conclusion We have developed the P-MSCs isolation and culture system, which lays a foundation for the establishment of P-MSCs bank in our hospital.