Correction: A single-molecule counting approach for convenient and ultrasensitive measurement of restriction digest efficiencies.

[This corrects the article DOI: 10.1371/journal.pone.

A single-molecule counting approach for convenient and ultrasensitive measurement of restriction digest efficiencies.

Restriction endonucleases play a central role in the microbial immune system against viruses and are widely used in DNA specific cleavage, which is called restriction digestion, for genetic engineering. Herein, we applied digital cell-free protein synthesis as an easy-to-use orthogonal readout means to assess the restriction digest efficiency, a new application of digital bioassays. The digital counting principle enabled an unprecedentedly sensitive trace analysis of undigested DNA at the single-molecule level in a PCR-free manner.

Establishment and characterization of a human thyroid carcinoma cell line (HOTHC) producing colony stimulating factor.

A cell line designated HOTHC was established from an anaplastic carcinoma (giant cell type) of the thyroid gland of 80-year-old woman. The HOTHC line grew rapidly in multilayer without contact inhibition, and more than 120 serial passages were made within 27 months. The cells were spindle or polygonal in shape and revealed neoplastic and pleomorphic features.

Establishment and characterization of a human malignant choroids plexus papilloma cell line (HIBCPP).

A cell line designated "HIBSPP" was established from a human malignant choroids plexus papilloma of 29-year-old Japanese woman. This line grew well without interruption for 3 years and was subcultivated over 70 times. The cells were spindle, oval, and polygonal in shape, and neoplastic and pleomorphic features, a jigsaw puzzle-like arrangement, multilayering and forming papillary structures without contact inhibition.

Establishment and characterization of human glioblastoma cell line (HUBT-n).

We succeeded in primary culture of 3 in 4 cases of glioblastomas. The long-term passage cultures were not done from the primary cultures of original tumor, but glioblastoma cell line (HUBT-n) was established from a xenograft of nude mouse. This line grew well without interruption for 4 years and was subcultivated over 120 times.

Establishment and characterization of human malignant lymphoma cell line derived from uterine cervix.

Human uterine cervical malignant lymphoma (B-cell type) was cultured and the cell line (HIUML) was newly established. The HIUML cells were round in shape and had a tendency to make floating clusters. The cells had a smooth surface or protrusion on the margin of the cytoplasm, and proliferate in floatation.

In vitro culture of various typed meningiomas and characterization of a human malignant meningioma cell line (HKBMM).

We placed on culture the 13 cases of meningiomas, succeeded in making a primary culture of 10 cases and maintained 5 cases in vitro over considerable period of time (over three month), and one cell line derived from a malignant meningioma were established. In the early period of the primary culture, meningioma cells were spindle- or round-shaped cells. In the case of psammomatous type, the cultured cells were characterized as forming psammoma bodies.

Establishment and characterization of human uterine cervical epidermoid carcinoma cell line HHUS containing HPV 59 DNA.

The cell line designated HHUS was established from human uterine cervical keratinizing squamous cell carcinoma. The HHUS cell line was subcultivated more than 70 times within 3 years. The cultured cells, polygonal or spindle, with neoplastic and pleomorphic features, appeared epithelial in shape, with a pavement-like arrangement and grew in multi-layers without contact inhibition.

Histogenesis of carcinosarcoma and establishment of leiomyosarcoma cell line (HTMMT) derived from human uterine carcinosarcoma.

A cell line designated HTMMT was established from the human uterine carcinosarcoma (composed of leiomyosarcoma and adenocarcinoma) of a 66-year-old Japanese woman. The cell line grew well and 83 serial passages were successively done within 24 months. The cell line contained spindle- or fibrous-shaped cells that revealed neoplastic and pleomorphic features, and multipled rapidly without contact inhibition.

Biological characteristics of cultured cells derived from various types of human brain tumors.

We placed in culture brain tumors from 45 cases (7 cases of astrocytoma, 2 from oligodendrogliomas, 2 glioblastomas, 2 ependymomas, 13 meningiomas, 6 pituitary adenomas, 5 neurinomas, a malignant lymphoma, a choroid plexus papilloma, and 6 metastatic tumors) and succeeded in making a primary culture from 33, and maintained 17 in vitro over a considerable period of time (greater than three months). In the early period of the primary cultures, the astrocytoma cells had cytoplasmic processes which contacted each other, the oligodendroglioma cells were small and spindle-shaped, the glioblastoma cells were neoplastic with pleopmorphic features and possessed cytoplasmic processes, the ependymoma cells formed a rosette-like cell arrangement, the meningioma cells were spindle- or round-shaped cells and characterized as forming psammoma bodies, the pituitary adenoma cells were round- or oval-shaped cells and produced growth hormone (GH), adenocorticoid tropic hormone (ACTH), prolactin, or other hypophyseal hormones, the choroid plexus papilloma cells were round-or polygonal and showed a papillary cell arrangement, the neurinoma cells were spindle- or fibrous-shaped cells, and the malignant lymphoma cells were round and formed cell aggregates floating in the culture medium.

Establishment and characterization of a human gestational choriocarcinoma cell line (HOCC).

Human gestational choriocarcinoma cell line (HOCC) was established from the mouse graft of choriocarcinoma. The HOCC cells were spindle or polygonal in shape and multi-nucleated giant cells, showing neoplastic and pleomorphic features. The cell proliferated stably, and the population doubling time was about 32 hours.

Establishment and characterization of a human malignant mesothelioma cell line (HMMME).

A cell line designated "HMMME" was established from the pleural fluids of a malignant mesothelioma patient. This line grew well without interruption for 12 years and was subcultured over 200 times. The cells were spindle and roundish in shape and displayed a monolayer sheet in an epithelial pavement cell arrangement.

Establishment and characterization of two squamous cell carcinoma cell lines (HYVC and HMVC) derived from vulva.

Two cell lines, HYVC and HMVC, were established from cultures of the squamous cell carcinoma removed from the vulva of females of 37 and 46 years old, respectively. These cell lines were very similar in their biological characteristics, such as morphology (polygonal), growth pattern (32-43 hr of population doubling time, 50-25% of plating efficiency), heterotransplantability (1 x 10(7) cells produced squamous cell carcinomas in the nude mice), genetics (75-78 of modal chromosomal number), and producing the tumor markers (SCC and TPA). The HPV-DNA was not detected in either the HYVC or HMVC lines by using L1-PCR methods, however, it was detected in the HYVC using E6/E7-PCR.

Establishment and characterization of a lymphoepithelial-like carcinoma cell line (HUUCLEC) derived from the human uterine cervix.

A cell line designated HUUCLEC was established from a human uterine cervical lymphoepithelial carcinoma obtained from a 61-year-old Japanese woman. The cell line has grown slowly without interruption and serial passages were successively carried out 60 times within 3 years. The cultured cells were spindle or round in shape, showing anaplastic and pleomorphic features, a pavement cell arrangement and multilayering without contact inhibition.