Vol.1 No.3 2009

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Research paper : Development of regenerative medical technology working toward practical application (H. Ohgushi)−156 Synthesiology - English edition Vol.1 No.3 (2009) While growing and processing cells are conducted in CPC, cells that were harvested from patients in hospitals contain various cells other than target cells. Target cells must be isolated and then grown. To accomplish this, development of cell selection technique and evaluation of whether the selected cells are growing well is necessary. The grown cells are converted to specialized cells after process of differentiation so they may develop into tissues or organs that must be regenerated. Evaluation of whether the differentiated cells possess ability as specialized cells is also necessary. In some cases the grown cells are transplanted directly to patient, but in many cases (for example, in our work of bone and joint regeneration), cells and biomaterials are combined and this hybrid material is transplanted. In this case, it is also necessary to evaluate safety of the biomaterial used as well as efficacy of the cells, such as whether the material supports cell differentiation. Standardizing these evaluation processes will enable application to greater number of patients, and hence the treatment may be accepted by the society. The above discussion can be summarized into four points.1) Providing environment for cell processing center (CPC)2) Selection of target cell and verification of proliferative ability3) Verification of cell differentiation (verification of biomaterial)4) Standardization of regenerative medicine3 Our efforts on regenerative medicine issues3.1 Providing environment for human cell processing center (CPC)Figure 1 shows the diagram of cultivation process. Cells (bone marrow) are harvested from the patient in hospital. The cells are transported to our CPC where they are grown. Although in some cases the grown cells are directly transplanted, they are differentiated into component cells of tissues or organs after further differentiation process. The differentiated cells are delivered to the hospital and transplanted to the patient in the hospital. Normally, this differentiation process takes place on various biomaterials[1].As mentioned earlier, when bacteria or mold contaminate the culturing processes, bacteria grow along with target cells and the cells become unsuitable for use. Bacteria are present in ordinary environment. Therefore, cell culture maneuvers are conducted in sterile cabinets of the CPC, which is supplied with air from which fine particles are removed by HEPA filter used in clean room in semiconductor plants. Even if sterile environment can be created physically in the CPC, various bacteria exist in human body, and human operator may serve as a source of contamination. However, an operator is absolutely necessary for growing and processing cells in CPC. Also, observation of cell through microscope in the CPC is necessary to check whether the cells are growing and differentiating properly. To minimize entry and exit of operators into the CPC, we developed an automatic cell observation device with Sanyo Electric Co., Ltd.[2]. As shown in Figure 2, using this device, image of any culture dish in any position designated by the user can be observed from remote location via LAN. The cells can be observed without entering the CPC, and sterile environment remains intact. Moreover, culture processing is conducted according to strict quality control procedures, and the work of recording cultured cell data increases the workload of the operator.Figure 3 shows the images of cell observed at every 24-hour interval using the device. All observations were made for same area (fixed point). The number of cells increases with passage of time, and one can see that the cells are growing properly. There had been no device that allowed remote observation of microscopic field of cell culture at regular time intervals with reproducible results, and thus the significance of this development is high. The device also allows regular recording of observation results as image data. We were able to develop technology that contributes to improvement of quality control and reduction of worker load. Ideally, cell culture that involves no human hands is desirable, and we are working on automatic cultivation device, but I shall not (2)−Other facilitiesOfficeObservation roomDeveloped by Sanyo Electric Co.ServerControl roomAutomatic observation deviceClean roomHuman Cell Processing Center (CPC)LAN24-hour interval observation of human cells at fixed-pointFig. 2 Providing environment for human cell culture (development of automatic cell observation device).Fig. 3 Fixed-point observation by automatic cell observation device (automatic transportation incubator with cell observation function).

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