Stem cell characteristics of human malignant melanoma sphere cells cultured in vitro

WEN Huicai; SUI Yunpeng; JIAN Xueping; LIAO Huaiwei; MA Li; XU Guizhen; LIU Yanping; WAN Jun

doi:10.3969/j.issn.1002-266X.2016.19.007

Abstract：Objective To observe the stem cell properties of human malignant melanoma cells cultured in serum-free me-dium.Methods Human malignant melanoma A375 cell line was cultured in serum-free medium, and the melanoma sphere cells (sphere group) and common adherent cells (adherent group) were obtained.Cell migration was detected using Transwell cham-ber.CD20 and CD133 were detected by immunofluorescence.Sixteen BALB/c mice were divided into the experimental group and control group (8 rats in each group), and the sphere cells and the adherent cells of 1 mL (104 ) were injected into the mouse shoulder blade by subcutaneous injection in the sphere and the adherent groups, respectively.The tumor cells were observed and compared between the two groups.Six weeks later, the mice were killed to obtain the tumor tissues.Results A375 cells were seeded in serum-free medium, visible small round ball suspension cells formed and gradually becomes larger , the ball inside the cells were closely connected with good refraction.The cell migration ability and the expression intensity of CD20 and CD133 in the sphere group were higher than those in the adherent group .After 6 weeks of injection, the mice in the experimental group had a large tumor, and no tumor growth was found in the control group.The tumor tissues of the experimental group were stained with HE, and the malignant melanoma was pathologically diagnosed.Conclusions Human malignant melanoma cells cultured in serum-free medium have the characteristics of stem cells with high migration ability and tumorigenesis .High expression of im-mune markers CD20, CD133 is found and malignant melanoma exists in the tumor stem cells.