cell engineering division - rikenen.brc.riken.jp/info/pdf/03annual/08.pdf · cell engineering...
TRANSCRIPT
37
"Cell Engineering Division in RIKEN BioResource Center (BRC)” has been originally
organized as the “RIKEN Cell Bank” in 1987. It is a unique, non-profitable public organization
for deposit, isolation, standardization, preservation, and distribution of cultured animal
cell lines produced by life science research community. The staffs conduct not only banking
of cell resources but also researches and developments related to cell engineering, such as
establishment of novel animal cell lines and development of technology to culture primate
embryonic stem cells efficiently.
1. Cell Banking: Animal cell lines
2. Cell Banking: Human somatic stem cells
3. Cell Banking: EBV transformed human B cells
4. Development of technology for cell banking
5. Development of technology to culture primate embryonic stem (ES) cells
6. Development of technology to obtain human feeder cells that maintain ES cells
7. Development of technology to establish animal cell lines possessing multi-potency
Senior Scientists, Heads of Cell Engineering Division
Yukio NAKAMURA, Ph.D. ( 2003. 4~ )
Tadao OHNO, Ph.D. (2001. 4~ 2003. 3)
Research Scientist
Takashi HIROYAMA, Ph.D. (2003. 7~ )
Senior Technical Scientists
Mariko NAGAYOSHI (2001. 4~ )
Kaoru SAIJO (2001. 4~ )
Technical Staffs
Tsuyoshi FUJIOKA (2003. 4~ )
BRC Technical Staffs
Lang HUANG (2001.4~ 2003.5) Hideki HARADA (2001.4~ 2003.3)
Takeshi OYAMATSU (2001.4~ 2003.3) Satoru WATANABE (2001.4~ 2003.3)
Goal
Activities
Members
Cell Engineering Division
Yukio NAKAMURA
Senior Scientist, Head of Cell Engineering Division (April 2003~ )
Tadao OHNO Senior Scientist, Head of Cell Engineering Division (~ March 2003)
38
Mutsumi SHIRAI (2001.4~ 2003.3) Emi IIMURA (2001. 4~ )
Mayako KATO (2001. 4~ 2003. 8 ) Naoko AOKI (2001. 4~ )
Saeri TOTSUKA (2001. 4~ ) Michiko KUSUMI(2001. 4~ )
Noriko TAKAI (2001. 4~ ) Youko KODAIRA (2001. 4~ )
Sachiko UCHIYAMA (2001. 4~ ) Kanae KURITA (2002. 12~ )
Takako EHARA (2003. 1~ ) Mariko NAKAMURA (2003. 3~ )
Hiroko HORIKIRI (2001. 4~ ) Michiru YAMANAKA (2003.10~ )
Naomi YUHARA ( 2003. 10~) Ayae HUKUZAWA (2003. 11~ )
Collaborative Scientists
Takeshi TODOROKI, Ph. D. (2001. 4~ ) Akinori OKI, Ph. D. (2001. 4~ )
Koji KAWAI, Ph. D. (2001. 4~ ) Eiji UCHIMURA (2001.4~ )
Kuang MING (2001.4~ 2003.7) Takehiro OIKAWA, Ph.D. (2001. 4~ )
Hiroshi MATSUI, Ph. D. (2001. 4~ )
Resarch Fellows and Student Trainees
Ken-ichi MIHARADA (2003. 4~ ) Rui MAEDA (2003. 4~ )
Yumiko NAKAMURA (2001. 4~ ) Milla GHOSH (2001. 4~ )
Terukazu ENAMI (2003. 4~ 2003. 6 ) Osamu SHIMOKAWA, M. D. (2003. 7~ )
Junpei UDO, M. D. (2003. 8~ )
Aoki, Kusumi, Nakamura, Uchiyama, Ehara, Kurita, Miharada, Kodaira,
Iimura, Nagayoshi, Horikiri, Maeda, Takai, Saijo, Yuhara,
Hiroyama, Nakamura, Totsuka
Dr. Ohno
(~March 2003)
��� ������ ��������������� ���� ����������� ��������
39
Specific aim 1. Cell Banking: Animal cell lines
We are possessing over 1,500 cell lines, of which 870 lines are available for distribution.
Recently nearly 3,000 ampoules have been distributed in a year, mostly to research
organizations (70%) and private organizations – approximately 10% overseas. We will
continue to accept donations of cultured animal cell lines and expand the collection, since the
significance of those cell lines in the field of biology will never diminish.
2. Cell Banking: Human somatic stem cells
Compared to obtaining primary cells derived from experimental animals it is quite difficult
to obtain human primary cells. The recent life science research, however, require human
primary cells, such as stem cells, especially in the fields of transplantation and regenerative
medicine. We are trying to establish a system to provide such human primary cells efficiently.
First, collaborating with ‘‘Clinical Cord Blood Bank’’ we are planning to supply human
cord blood that is not useful for transplantation to domestic researchers in order to contribute
for the fields of transplantation and regenerative medicine. Cord blood is a source of not only
hematopoietic stem cells but also other somatic stem cells.
Second, collaborating with researchers who developed technologies to expand human
mesenchymal stem cells in vitro very efficiently we are planning to supply human
mesenchymal stem cells to researchers. Mesenchymal stem cells can differentiate to bone and
cartilage cells and reportedly also to neuron and cardiomyocyte.
3. Cell Banking: EBV transformed human B cells
In order to analyze the causes of certain specific diseases in the level of genome, a lot of
genome samples are required. However, it is not so easy to collect such abundant samples for
each researcher. Thus the collection of a lot of genome samples and/or cell lines containing
the genome is very important and useful for researchers in the field. We are planning to collect
human B cell lines immortalized by Epstein-Barr virus transformation. The donator of B cells is
not only volunteers possessing certain disease but also healthy volunteers.
4. Development of technology for cell banking
Basically we are always challenging to improve technology for cell banking, such as culture
method, stock method, and control of information.
In order to maintain the quality of human cell resources we established good
manufacturing practice (GMP) facility this year (2003). We are planning to culture human
hematopoietic stem cells in cord blood and human mesenchymal stem cells in the facility.
Cell banking of human cells require strict regulation about ethical matters. We only accept
the donation of human cells that are admitted by the RIKEN Tsukuba Institutional Review
Board (IRB). Furthermore, RIKEN BRC contracts material transfer agreement (MTA) with the
organization that donates human cells to RIKEN BRC. In the MTA RIKEN BRC confirms that the human resource was obtained after a strict informed consent. An admission by the IRB in
��� ������ ��������������� ���� ����������� ��������
40
the organization that donates human cells to RIKEN BRC is also necessary. When RIKEN BRC
supplies human cells to user, RIKEN BRC contracts MTA with the organization that the user is
belonging. As for certain human cells, such as cord blood, an admission by the IRB in the user
organization is also required.
5. Development of technology to culture primate embryonic stem (ES) cells
Embryonic stem cells have a lot of potentials not only in the field of basic biology but
also in the field of clinical science. Before human ES cells are applied to clinical science, a lot
of experiments using primate ES cells should be required naturally. Then, we are planning a
banking of primate ES cells. At the moment we are acquiring the technology to culture primate
ES cells using a cynomolgus monkey ES cell line.
6. Development of technology to obtain human feeder cells that maintain ES cells
To our knowledge all human ES cell lines existing at the moment are cultured on the feeder
layer derived from mouse embryo, i.e. mouse embryonic fibroblast (MEF). Considering the
application of human ES cells to clinical science the cells cultured on MEF are not appropriate.
Feeder cells derived from human resources are necessary and required, however the
technology to obtain human feeder cells constantly is not established yet. We are planning to
establish such technology using fibroblastic cells derived from umbilical cord.
7. Development of technology to establish animal cell lines possessing multi-potency
Animal cell lines possessing multi-potency and/or tissue-specific character are very
useful for developmental biology and the basic research of regenerative medicine. We are
trying to establish such cell lines by various approaches. First, identification and purification
of tissue-specific stem cell may lead to establishment of such cell line by immortalizing the
stem cell. Second, induction of ES cell’s differentiation may lead to such cell line. Third,
reprogramming of somatic cells may lead to such cell line. We are investigating these
possibilities.
��� ������ ��������������� ���� ����������� ��������
41
Figure 1.Results of distribution of cell resources for these approximately 10 years. Nearly 3,000 ampoules are distributed to researchers every year recently.
Figure 2.Scheme of banking of human cord blood. Human cord blood that is originally donated for transplantation but is not suitable for the purpose will be supplied to researchers depending on the informed consent of donators.
��� ������ ��������������� ���� ����������� ��������
42
Original Papers (* Peer reviewed Journal)
1. Chen Y. Z., Hayashi Y., Wu J. G., Takaoka E., Soeda E. and et al.: "A BAC-Based
STS-Content Map Spanning a 35-Mb Region of Human Chromosome 1p35-p36",
Genomics 74: 55-70 (2001)*
2. Masters J. R., Thomson J. A., Daly-Burns B., Reid Y. A., Ohno T. and et al.: "Short tandem
repeat profiling provides an international reference standard for human cell lines", Proc.
Nat. Acad. Sci. USA 98: 8012-8017 (2001)*
3. Chen Y. Z., Soeda E., Yang H. W., Takita J., Chai L. and et al.: "Homozygous Deletion
in a Neuroblastoma Cell Line Defined by a High-Density STS Map Spanning Human
Chromosome Band 1p36", Genes, Chromosomes & Cancer 31: 326-332 (2001)*
4. Yang H. W., Chen Y. Z., Takita J., Soeda E., Piao H. Y. and Hayashi Y.: "Genomic structure
and mutational analysis of the human KIF1B gene which is homozygously deleted
inneuroblastoma at chromosome 1P36.2”, Oncogene 20: 5075-5083 (2001)*
Figure 3.Usefulness of human mesenchymal stem cells. Human mesenchymal stem cells that are originally donated for regenerative medicine etc. and expanded in vivo will be supplied to researchers depending on the informed consent of patients.
Publications
��� ������ ��������������� ���� ����������� ��������
43
5. Yang H. W., Chen Y. Z., Piao H. Y., Takita J., Soeda E. and Hayashi Y.: “DNA
Fragmentation Factor 45 (DFF45) Gene at 1p36.2 Is Homozygously Deleted and Encodes
Variant Transcripts in Neuroblastoma Cell Line”, Neoplasia 3: 165-169 (2001)*
6. H. Harada, K. Saijo, S. Watanabe, K. Tsuboi, T. Nose, I. Ishiwata, and T. Ohno.: “Selective
expansion of human natural killer cells from peripheral blood mononuclear cells by the
cell line, HFWT”, Jpn. J. Cancer Res. 93: 313-319 (2002)*
7. Bao Gang Peng, Shu Qin Liu, Ming Kuang, Qiang He, S. Totsuka, Jiefu Huang, Ming-De
Lu, Li-Jiang Liang, Kam W. Leong, and T. Ohno.: “Autologous fixed tumor vaccine: A
formulation with cytokine-microparticles for protective immunity against recurrence of
human hepatocellular carcinoma”, Jpn. J. Cancer Res. 93: 363-368 (2002)*
8. Nakamura, Y., Grumont, R.J., and Gerondakis, S.: “NF-kB1 can inhibit v-Abl-induced
lymphoid transformation by functioning as a negative regulator of cyclin D1 expression”
, Mol. Cell. Biol. 22: 5563-5574 (2002)*
9. Osawa, M., Yamaguchi, T., Nakamura, Y., Kaneko, S., Onodera, M., Sawada, K., Jegalian,
A., Wu, H., Nakauchi, H., and Iwama, A.: “Erythroid expansion mediated by the Gfi-1B
zinc finger protein: role in normal hematopoiesis”, Blood 100: 2769-2777 (2002)*
10. Fujiki, Y., Fukawa, K., Kameyama, K., Kudo, O., Onodera, M., Nakamura, Y.,
Yagami, K-i., Shiina, Y., Hamada, H., Shibuya, A., and Nakauchi, H.: “Successful
xenotransplantation of human hematopoietic progenitor cells into pig”, Transplantation
75: 916-922 (2003)*
OralPresentations
1. Ohno, T.: "Selective expansion of human natural killer cells and its application to brain
tumor therapy", 3rd symposium of Cell Biotechnology: practice and strategy of new
biotech era (2001).
2. Harada, H., Watanabe, S., Saijo, K., Ishiwata, I., and Ohno, T.: “Identification of
pre-natural killer (NK) cell populations specifically activated by a Wilm’s tumor cell
line”, American Association for Cancer Research (2002)
3. Emura, F., Koike, N., Kawamoto, T., Ghosh, M., Saijo, K., Ohno, T., and Todoroki, T.:
“Establishment and characterization of novel biliary tract cancer xenograft model”,
World Congress of the Hepatobiliary Pancreatic Association (2002)
��� ������ ��������������� ���� ����������� ��������
44
4. Yukio NAKAMURA: “Banking of human and animal cells: National BioResource Project”,
The 26th Annual Meeting of The Molecular Biology Society of Japan, Kobe (2003)
5. Takashi HIROYAMA, Toyoki MAEDA, Yukoh NAKAJIMA, Ryo KURITA, Kazumi ODA,
Rie ARIYOSHI, Kenichi MIHARADA, Tsuyoshi FUJIOKA, Rui MAEDA, Yukio
NAKAMURA, Toshitaka MUTOH, Tatsuya KAWAGUCHI, Hideki NAKAKUMA,
Taroh KINOSHITA, Kenzaburou TANI: “Suppression of megakaryocytic differentiation
of GPI-anchored protein defficient K562 cells and recovery from the suppression by
Musashi1.”, The 26th Annual Meeting of The Molecular Biology Society of Japan, Kobe
(2003)
6. Rui MAEDA, Kenichi MIHARADA, Takashi HIROYAMA, Tsuyoshi FUJIOKA, Kenichi
YAGAMI, Yukio NAKAMURA: “Functional analysis of D type Cyclin in hematopoiesis”,
The 26th Annual Meeting of The Molecular Biology Society of Japan, Kobe (2003)
7. Kenichi MIHARADA, Mitsujiro OSAWA, Takashi HIROYAMA, Tsuyoshi FUJIOKA, Rui
MAEDA, Toshiro NAGASAWA, Yukio NAKAMURA: “Role of 24p3, a novel transporter
of iron, in hematopoiesis”, The 26th Annual Meeting of The Molecular Biology Society of
Japan, Kobe (2003)
��� ������ ��������������� ���� ����������� ��������