| Authors |
Yoichi Kobayashi, Issei Kiguchi, Isamu Ishiwata, Bunpei Ishizuka, Megumi Iguchi, Yuko Tokieda, Chieko Ishiwata, Hiroshi Ishikawa; |
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| Abstract |
A cell line designated HNROA was established from the human uterine
cervical adenocarcinoma. The diagnosis made was Peutz-Jeghers syndrome
with uterine cervical adenocarcinoma. On September 2, 1999, a primary
uterine tumor was removed from a 53 year-old Japanese woman undergoing
radical hysterectomy and lymph node resection. The portion of tumor was
placed into culture the next day. Histopathological diagnosis of the tumor
indicated a minimal deviation adenocarcinoma (so-called adenoma malignum).
The material was minced with a sharp pair of scissors, treated with 600
Pronase units/ml dispase, and centrifuged. The tissues were placed in 6 cm
plastic dishes containing the mixed culture media of Ham's F12 and RPMI
1640 (GIBCO), 100 units of penicillin and 100 mug of streptomycin per ml,
and 15% fetal calf serum; the final pH was 7.2-7.4. The collagen-coated
dishes were incubated at 37 Celsius in a CO2 incubator supplied with
humidified 5% CO2 and 95% air. In the initiation of the primary culture,
an epithelial colony and fibroblastic cells appeared. Pure epithelial
cells were obtained by the isolation technique devised by Okumura. The
cell line has grown slowly and serial passages were successively carried
out 21 times within 9 months. The cultured cells were spindle, round, and
columnar in shape, showing anaplastic and pleomorphic features and
multilayering without contact inhibition. The cells formed gland-like
structures in vitro. The population doubling time was 83 hours. The
chromosomes showed a wide distribution of aneuploidy, the mode was in the
hypo-triploid range, and many marker chromosomes were observed. The cells
produced a large amount of CEA and TPA but were not able to be transplanted
into the nude mouse.
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