Characterization of a human ovarian teratocarcinoma‐derived cell line
Authors: Zeuthen J. , Nørgaard J. O. R. , Avner P. , Fellous M. , Wartiovaara J. , Vaheri A. , Rosén A. , Giovanella B. C. 
Published: February 2007

A cell line (PA I), derived from human ovarian teratocarcinoma cells, was obtained by culturing ascitic fluid cells from a patient with recurrence of malignant ovarian teratoma. During early passages the cultured cells showed a variable morphology, a long doubling time, and a low plating efficiency (2 %). After about 50 passages in vitro, a cell population which was more homogenous and resembled embryonal carcinoma cells were obtained. These cells had a shorter doubling time (26 h), and increased plating efficiency (77 %). The early‐passage cells were aneuploid (P 24) whereas the late‐passage cells had a normal diploid karyotype with one balanced translocation between chromosomes No. 15 and No. 20 (P 224). Details of the karyotype suggest that the cells are heterozygous, i.e. derived from a stage before the first meiotic division. One of the two X chromosomes was inactive, and the cells expressed HLA antigens (A28 and B12), and β2‐microglobulin. Expression of F9 antigen, characteristic of two‐cell and later preimplantation embryos, was absent, while expression of PCC4 antigen, expressed also by blastocysts, was present. This finding suggests that the line might express some embryonic characteristics.

The PA I cell line maintained in monolayer cultures showed several characteristics of malignant cells. The proportion of malignant cells increased with successive passages in vitro. The late‐passage cells represented a fairly homogenous population of malignant cells similar to embryonal carcinoma cells. Late‐passage PA I cells, when seeded under conditions that prevented attachment of cells to the substratum, formed embryoid bodies consisting of an inner core of cells similar to embryonal carcinoma cells, surrounded by a rind of endoderm‐like cells. These two cell layers were separated by a basement membrane‐like structure containing fibronectin. The core embryonal carcinoma cells expressed high alkaline phosphatase activity whereas the endoderm‐like cells had low alkaline phosphatase activity. Embryoid bodies seeded on an adhesive substratum formed polycystic structures divided by layers of epithe‐lial‐like cells and containing extracellular fibrils similar to collagen type I or III. In these cultures, further limited differentiation into endoderm‐like, epithelial‐like cells and pigmented cells was observed. Morphological differentiation of undifferentiated PA I cells into endoderm‐like cells in monolayer cultures could be obtained by treatment with BrdUrd or by plating in low serum concentration and at low density. Cells with characteristic fibrillar distribution of fibronectin and actin microfilament bundles were then observed, indicating formation of cells lacking properties of malignant cells. As indicated by these results, the PA I cell line, in spite of a limited capacity to differentiate in vitro, shares some of the properties of mouse teratocarcinoma cell lines and might therefore serve as a useful model for studies on some developmental mechanisms in human cells.