[Transformation of attached cells derived from fetal umbilical cord blood induced by conditional medium of hepatoma cells].
OBJECTIVE
Stem cells derived from fetal umbilical cord blood are of undifferentiated at early stage. They are sensitive to stimulations from the environment, and may be transformed under the effects of carcinogenic factors. This study was to explore the sensitivity of stem cells derived from fetal umbilical cord blood to carcinogenic factors.


METHODS
Mononuclear cells were isolated from fetal umbilical cord blood, and the attached cells were cultured in the medium containing 10% conditional medium of HepG2 hepatoma cells. A new cell line was gained, termed H-UCB. The biological features of H-UCB cells were detected by electron microscopy, karyotype analysis, cell cytometry, Western blot, and colony formation assay.


RESULTS
H-UCB cells proliferated faster after passage 3. The cells were fibroblast-like and hepatocyte-like, with the ratio of nucleus to cytoplasm increased. Under electron microscope, many microvilli on the surface and numbers of vacuoles in the cytoplasm of the cells were observed, the nuclei were large and irregular, endocytosis phenomena were noticed. Karyotype analysis indicated that the cells were heteroploid, and the number of chromosomes was between 50 and 70. Flow cytometry data indicated that the proliferation period was 22.9 h, and the karyotype was between diploid and tetraploid. Western blot showed that c-Myc protein and proliferating cell nuclear antigen (PCNA) were overexpressed in H-UCB cells. According to flow cytometry, the positive rates of surface markers of H-UCB cells were 79.0% for CD105, 1.2% for CD34, and 12.2% for CD106; those of control HepG2 cells were 15.0% for CO105, 9.8% for CD34, and 1.4% for CD106. The colony formation rate of H-UCB cells in soft agar was (13.2+/-2.6)%.


CONCLUSIONS
H-UCB cells are derived from endothelial cells, and are transformed as malignant cells with tumor cell characteristics.
Zhang
Xiao-Dong
XD
College of Life Sciences, Nankai University, Tianjin 300071, P. R. China. zhangxd@nankai.edu.cn
Cai
Na
N
Wang
Hong-Hui
HH
Guo
Shi-Yi
SY
Ye
Li-Hong
LH
D015703:Antigens, CD / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002461:Cell Line, Transformed / Q000378:metabolism / Q000648:ultrastructure; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D000071063:Endoglin; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D011123:Polyploidy; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
D015703:Antigens, CD; D018952:Antigens, CD34; D017077:Culture Media, Conditioned; C064206:ENG protein, human; D000071063:Endoglin; D018809:Proliferating Cell Nuclear Antigen; D016271:Proto-Oncogene Proteins c-myc; D011956:Receptors, Cell Surface; D019010:Vascular Cell Adhesion Molecule-1