Objective To investigate phenotypes molecules of macrophage derived from human pluripotent stem cell (hPSC), umbilical cord blood CD34+ hematopoietic stem/progenitor cell and leukemia cell line THP-1 and inflammatory cytokine mRNA relative expression levels under different polarization states, exploring functional differences of macrophage from these 3 kinds of sources. Methods Aforementioned 3 kinds of sources of macrophages, included macrophages derived from hPSC cultured with AGM-S3 stromal cell, macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell, and macrophages derived from THP-1 were selected as study subjects. Researches on these 3 kinds of sources of macrophages included: ①Morphology observation and May-Gr Giemsa (MGG) staining were performed to determine whether macrophages were obtained. ②Phenotypes molecules were detected by flow cytometry. ③Expression levels of various inflammatory cytokines from 3 kinds of sources of macrophages under stimulation of lipopolysaccharide, interleukin (IL)-4 were detected by fluorescent quantitative PCR, and different polarized states were confirmed. Independent sample t test was used to compare the relative expression levels of various inflammatory cytokines mRNA under different stimuli from 3 kinds of sources of macrophages. Results ① According to cell morphological observation and results of MGG staining, 3 kinds of sources of macrophages were successfully obtained. ② Results of flow cytometry showed, all 3 kinds of sources of macrophages expressed CD45, human leukocyte antigen (HLA)-Ⅱ, CD36 and CD11b. Macrophages derived from THP-1 only expressed macrophage phenotypes related molecules CD64 and CD11c. Macrophages derived from hPSC cultured with AGM-S3 stromal cell and macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell expressed macrophage phenotypes related molecules CD14, CD64, CD11c, CD68, CD86, CD206. ③ Results of fluorescent quantitative PCR showed, mRNA relative expression levels of IL-1β, -6, -12β, tumor necrosis factor (TNF)-α from macrophages derived from hPSC cultured with AGM-S3 stromal cell which peaked after 3 h of lipopolysaccharide stimulation, were (14.69±0.24), (305.50±67.26), (10.91±1.48), (128.31±9.90), and were higher than those under IL-4 stimulation, which were (0.75±0.14), (2.66±0.27), (0.54±0.04), (0.89±0.08) respectively, and the differences were statistically significant (t=85.630, 7.798, 12.128, 22.295; P=0.000, 0.001, 0.000, 0.000). mRNA relative expression levels of IL-1β, -6 from macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell which peaked after 9 h of lipopolysaccharide stimulation, were (16.80±0.56), (481.50±26.81), and were higher than those under IL-4 stimulation, which were (0.02±0.00), (0.51±0.23) respectively, and the differences were statistically significant (t=52.133, 31.080; P=0.000, 0.000). While mRNA relative expression levels of IL-12β, TNF-α which peaked after 3 h of lipopolysaccharide stimulation, were (56.48±3.78), (11.12±0.76), and were higher than those under IL-4 stimulation, which were (3.53±0.46), (0.33±0.18) respectively, and the differences were statistically significant (t=24.090, 24.010; P=0.000, 0.000). mRNA relative expression levels of TNF-α from macrophages derived from THP-1 which peaked after 3 h of lipopolysaccharide stimulation, was (21 388.05±2 464.90), and was higher than that under IL-4 stimulation, which was (143.89±5.30), and the difference was statistically significant (t=14.930, P=0.000). While mRNA relative expression levels of IL-1β, -6, -12β which peaked after 9 h of lipopolysaccharide stimulation, were (4.33±0.01), (14.53±3.79), (525.56±34.65), and were higher than those under IL-4 stimulation, which were (0.48±0.08), (0.01±0.00), (1.03±0.26) respectively, and the differences were statistically significant (t=82.710, 6.640, 26.220; P=0.000, 0.003, 0.000). Under stimulation of lipopolysaccharide, 3 kinds of sources of macrophages could effectively be polarized into type M1 macrophages. After 9 h of IL-4 stimulation, mRNA relative expression levels of IL-10 from macrophages derived from THP-1 was (262.79±20.18), and was higher than that under lipopolysaccharide stimulation, which was (1.71±0.02), and the difference was statistically significant (t=22.410, P=0.000). Under stimulation of IL-4, only macrophages derived from THP-1 could be effectively polarized into type M2 macrophages, and macrophages derived from hPSC cultured with AGM-S3 stromal cell and macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell both couldn′t be polarized into type M2 macrophages. Conclusions There were significant differences in phenotypes molecules expression on macrophages from three different kinds of sources. Macrophages derived from hPSC cultured with AGM-S3 stromal cell and macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell expressed more macrophage phenotypes related molecules. Macrophages derived from THP-1 lacked some macrophage phenotypes related molecules, and it was different from macrophage in vivo. Macrophages derived from hPSC cultured with AGM-S3 stromal cell and macrophages derived from umbilical cord blood CD34+ hematopoietic stem/progenitor cell could be polarized into type M1 macrophages in a short time of stimulation, but not type M2 macrophages. Macrophages derived from THP-1 could be polarized into both type M1 and type M2 macrophages in a long time of stimulation. Key words: Macrophages; Human pluripotent stem cells; Human embryonic stem cells; Fetal blood; Antigens, surface; Cytokines