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)

��� ������ ��������������� ���� ����������� ��������