Human pluripotent stem (ES/iPS) cells
Proliferation Control Medium
R&D; Bourbon Biomedical Advanced Research Laboratories, Inc.
Supplier; Nipro Corporation

72hr change-free medium culture ofhuman ES/iPS cells in Xyltech BOF-01
BOF-01 is a novel basal culture medium which can suppress proliferation of human
ES/iPS cells on feeder cell layers.
●BOF-01 can be used by completely replacing the basal culture medium(e.g. DMEM/F12) of the human ES/iPS cells.
●Human ES/iPS cells can be maintained with BOF-01 for about 3days (up to 72 hours)
without changing medium under normal culture conditions (37°C, 5% CO2).
●After suppression of cell proliferation with BOF-01, cell growth can be resumed bychanging normal growth medium.
●This product does not contain glucose.
●Note・It cannot be used under feeder-free culture conditions.
・BOF-01 does not contained growth factors nor supplements (such as KSR, bFGF etc.).It is required toadd necessary factors appropriately to prepare complete culture medium.
・Not all human ES/iPS cell lines have been tested with this product.
None of non-stem cell lines are proﬁled either.
・This product does not inhibit cell proliferation completely.
・Cellular condition under BOF-01 culture depends on the state and culture condition of the human ES/iPS cells.
・This product is for research use only, and not permitted for human or animal diagnostic or therapeutic uses.

Overview of Weekend-free Protocol●Cell proliferation control by BOF-01 made it possible to maintain/preserve cells during
the weekend in a normal culture environment without changing the medium.
Cell Passaging

Week 1MONTUEMON

Cell Passaging withBOF-01 mediumTHUFRISATSUN

Maintain cell culture, experiments, repeat cell passaging schedule with BOF-01 medium on Friday

Back tonormal growth medium

Week 2WED

Cell maintenance withproliferation control(No medium change)

TUE

WEDTHUFRISATSUN

The timing of passage needs to be adjusted to the cellʼ s condition and growth rate.

Morphology of Undiﬀerentiated Cell Colony and Eﬀect of Proliferation Control●Phase images of cell colonies and alkaline phosphatase stain of human iPS cells
before and after proliferation control culture by BOF-01 (201B7 strain).

●Comparison of cell proliferation rates of hiPS cells cultured in BOF-01 medium
and normal human ES/iPS growth medium.

Average fold expansion

8Normal culture

BOF-01 medium

Human ES/iPS growth medium

6Xyltech BOF-01 medium
4200123Days

456※The composition of normal growth medium (human ES/iPS medium)and BOF-01 medium are based on reference.
Curr Protoc Stem Cell Biol.2009 Jun; Chapter 4: Unit 4A.2.

Xyltech BOF-01 suppressed cell proliferation while maintaining human iPS cells in undiﬀerentiated state.

Undiﬀerentiation markers expression in human iPS cells by BOF-01●The high expression of pluripotent stem cell markers was conﬁrmed by immunoﬂuorescencestaining of human iPS cells (201B7) maintained in BOF-01 medium for 3 days.

Oct3/4Sox2

DAPI

Xyltech BOF-01 maintained human iPS cells in undiﬀerentiated state.

Diﬀerentiation of Three Germ Layers (Teratoma Assay)●The ability of diﬀerentiation was conﬁrmed by transplanting into immunodeﬁcient mouse byhuman iPS cells (201B7) maintained in BOF-01 medium for 3 days to form teratoma.

Mesoderm(Chondrocyte)

Endoderm(Alimentary canal epitherial cell)

Ectoderm(Pigment cell)

Xyltech BOF-01 maintained human iPS cells in pluripotent state.

Colony morphology changes during proliferation control culture
HumanES/iPS cells maychange the morphology ofcolonies while cultured in
BOF-01 medium. The originalmorphology can be resumed
after changing back tonormal human ES/iPS
medium (Phase images show
the example of 201B7 cells).

Normal culture (Day 1)

BOF-01 medium (Day 4)

Back to normal culture (Day 5)

Code No.

Product TypeExpiration

StorageSize

87 ー 280

Xyltech BOF-01

12 months

２〜８℃

100mLBourbon Biomedical Advanced
Research Laboratories, Inc.

1-3-1, Ekimae, Kashiwazaki City, Niigata Pref.,945-0055, Japan
E-mail: support@bourbon-barl.co.jp