2004 Fiscal Year Final Research Report Summary
Construction of human cell culture system for regenerative medicine without using animal components
| Project/Area Number |
14380406
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | RIKEN (The Institute of Physical and Chemical Research) (2004)
Kanagawa Academy of Science and Technology (2002-2003) |
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Principal Investigator |
ITO Yoshihiro RIKEN, Nano Medical Eng.Lab., Chief Scientist, 伊藤ナノ医工学研究室, 主任研究員 (40192497)
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| Co-Investigator(Kenkyū-buntansha) |
NOGAWA Masayuki Kanagawa Academy of Science and Technology, Regenerative Medical Reactor Project, Researcher, 再生医療バイオリアクタープロジェクト, 研究員 (90359117)
IKEBUCHI Kenji Saitama Medical University, Department of Medicine, Professor, 医学部, 教授 (20175194)
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| Project Period (FY) |
2002 – 2004
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| Keywords | Cord Blood / Hematopoietic Stem Cell / CD34 Positive Cell / Cytokine / Cell Immobilization / Nurse Cell / Embryonic Stem Cell / Artificial Cell Culture Matrix |
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| Research Abstract |
It is very important to expand the number of human hematopoietie stem cells in cord blood without differentiation, because the cell number is not enough for patients of heavy weight. In addition, in the case human embryonic stem cell there is no way to culture without murine fibroblast cells. Therefore, we have tried to develop culture systems of human stem cells without using animal feeder cells.
1.New human bone marrow cells incorporated with telomerase gene were established with cooperation with Cancer Research Center. Some cell lines were cloned and the clones supported growth of hematopoietie stem cells(HSC) in cord blood. However, in these cases, the growth of cell line was too fast to isolate HSC from the cell line. Therefore, the cell line was chemically fixed and employed for supporting growth of HSC.
2.When some cells supporting growth of HSC were induced to fat cells, the supporting abilities disappeared. Therefore, the difference of proteins before and after the differentiation was investigated and some differences were observed by two dimensional electrophoresis. Now the gene expression is also investigated by a DNA microarray. When the protein or gene for supporting growth ofHSC, the protein will be immobilized on a substrate for artificial cell culture matrix.
3.Monkey embryonic stem(ES) cell was cultured on human placenta feeder layers without differentiation of ES cells. Considering that he placenta was usually discarded as a medical waste, the success is very useful. Although the monkey ES cell was used as a preclinical research, the similarity can be extended to human ES cells and it will be very useful for practical regenerative medicine.
4.Mouse ES cells were cultured on various artificial substrata and the growth was evaluated. It was found that chemically fixed feeder cell supported the growth of murine ES cells. Now this fixation is applied for placenta for culture of monkey ES cells.
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Research Products
(14 results)
