Fabrication of tissue-engineered cell sheets by automated cell culture equipment.

Most cells for regenerative medicine are currently cultured manually. In order to promote the widespread use of regenerative medicine, it will be necessary to develop automated culture techniques so that cells can be produced in greater quantities at lower cost and with more stable quality. In the field of regenerative medicine technology, cell sheet therapy is an effective tissue engineering technique whereby cells can be grafted by attaching them to a target site. We have developed automated cell culture equipment to promote the use of this cell sheet regenerative treatment. This equipment features a fully closed culture vessel and circuit system that avoids contamination with bacteria and the like from the external environment, and it was designed to allow 10 cell sheets to be simultaneously cultured in parallel. We used this equipment to fabricate 50 sheets of human oral mucosal epithelial cells in five automated culture tests in this trial. By analyzing these sheets, we confirmed that 49 of the 50 sheets satisfied the quality standards of clinical research. To compare the characteristics of automatically fabricated cell sheets with those of manually fabricated cell sheets, we performed histological analyses using immunostaining and transmission electron microscopy. The results confirmed that cell sheets fabricated with the automated cell culture are differentiated in the same way as cultures fabricated manually.

Real-time explosive particle detection using a cyclone particle concentrator.

RATIONALE: There is a need for more rapid methods for the detection of explosive particles. We have developed a novel real-time analysis technique for explosive particles that uses a cyclone particle concentrator. This technique can analyze sample surfaces for the presence of particles from explosives such as TNT and RDX within 3 s, which is much faster than is possible by conventional methods. METHODS: Particles are detached from the sample surface with air jet pulses, and then introduced into a cyclone particle concentrator with a high pumping speed of about 80 L/min. A vaporizer placed at the bottom of the cyclone particle concentrator immediately converts the particles into a vapor. The vapor is then ionized in the atmospheric pressure chemical ionization (APCI) source of a linear ion trap mass spectrometer. RESULTS: An online connection between the vaporizer and a mass spectrometer enables high-speed detection within a few seconds, compared with the conventional off-line heating method that takes more than 10 s to raise the temperature of a sample filter unit. Since the configuration enriched the number density of explosive particles by about 80 times compared with that without the concentrator, a sub-ng amount of TNT particles on a surface was detectable. CONCLUSIONS: The detection limit of our technique is comparable with that of an explosives trace detector using ion mobility spectrometry. The technique will be beneficial for trace detection in security applications, because it detects explosive particles on the surface more speedily than conventional methods.

Intradural cranial chordoma.

BACKGROUND: Intradural chordomas are rare and have been considered benign owing to the feasibility of complete resection and the display of lesser aggressive biologic behavior than typical chordomas. CASE DESCRIPTION: We herein reported 2 cases of intradural cranial chordoma with aggressive biologic behavior. A tumor (anti-Ki-67 monoclonal antibody [MIB-1], 13.9%) in a 59-year-old woman was strongly adherent to the brainstem and involved the basilar artery and its branches. After subtotal removal, the remnant tumor was treated with stereotactic radiotherapy. A tumor (MIB-1, 6.2%) in a 75-year-old woman repeatedly recurred even after initial gross total removal. The recurrent chordomas were treated with γ-knife radiosurgery. CONCLUSION: The cases presented in this study indicate that intradural chordomas can also be aggressive such as typical chordomas. Long-term follow-ups with a large number of patients with this condition are essential for elucidating the prognosis of intradural chordomas.