Evaluation based on Alternative Method to Animal Testing

Safety evaluation
contract service

Bringing innovation to safety evaluation with accurate blood cells andnew test methods of measuring command substances.
In Alternative Methods to Animal Testing, there are many challenges based on
the differences between the cells used* and normal human cells. MiCAN has
solved this problem by using a brand-new approach based on regenerative
medicine technologies and drastically increased the reliability of evaluation

results and choice of evaluation designs. We are offering essential evaluations
for the development of pharmaceutical products and cosmetics at low cost inan efficient and speedy manner. Our cutting-edge technology will drasticallychange the future development of pharmaceutical products and cosmetics.
* Cell lines represented by THP-1, Mono-Mac-6, etc.

Highly accurate examination is now available with Mylc, new blood cells for tests.
Bloodcells

Mylc blood cells are produced from healthy human cells using regenerative medicine technologies. Mylc blood cells, which arecontrolled at a pre-stimulated (immature) state before receiving external stimuli and are genetically homogeneous,show pure and clear responses to stimulations in examinations and assure highly reliable test results.
Mylc blood cells show very similar biological responses to those of humans and have no problems in reproducibility of results.

Differentiation and maturation of dendritic cells
iPS/ES cell

CFU-GM

CFU-M

MonoblastDendritic cell
( Immature )White blood cell
( Monocytes )Dendritic cell
( Antigen presenting cell )Stimulation

Mylc

Can be producedfrom conventional humanstem cells.

Optimum differentiation
state for evaluation

T cellTransmitter

Release transmitter
responding to stimulation

Macrophage

Charac teris tics o f M ylc
Homogeneous blood cellswith identical genetic information
We can control the differentiation stage
to the optimum conditions
Stable mass-production
and supply are possible

The same genetic information and conditions can be always reproduced because they are
produced in our laboratory. This feature contributes to a drastic improvement in the accuracy andapplicability of pharmaceutical evaluation.
Differentiation stage (maturity ) can be minutely controlled with our high level ofregenerative medicine technologies and product control.
Research and evaluation of high accuracy are available.
You can obtain blood cells in the sufficient quantities in ideal condition any time.
You can conduct research and development and efficacy evaluation with no delay andaccelerate your development.

Three types of blood cells based on the origins.
Different genetic background can be selected depending on the purpose.
We provide three types of ( immature ) dendritic cells based on the original cell types. For example, tests examining individual characteristics
such as sensitive skin and allergic predisposition can be conducted, allowing a wide variety of evaluation designs.

aMylc
Human peripheral blood mononuclear cells
Blood cells that show in vivo-like characteristics andreactions.

iMylc
iPS cell-derived blood cells
Blood cells with standardized genetic background.
Standard reactions can be elicited.

uMylc
Customization using provided blood
Blood cell materials with characteristics of the donor
can be prepared.

Wide variety of evaluation designs are available• Want to compare the degrees of responses to drugs in humans with sensitive skin.
• Want to accurately determine response patterns in people who have idiosyncrasies.
• Want to advance development of drugs quickly by measuring drug effect in each evaluation. • Want to accurately evaluate and compare the effect of drugs under identical conditions.

TestmethodTest method

Our new test methods can greatly reduce the time and cost.
Major alternative methods to animal testing such as the h-CLAT, a skinsensitization assay, or a pyrogen test using cell lines have various
problems such as high cost, long evaluation time, reproducibility, andsensitivity ( in particular, false positives). On the other hand, in MiCAN’s
new method, which uses only iMylc-2 ( iPS cell-derived non-stimulated
dendritic cells ), the relationship between stimulation with substances
and the released transmitter ( IL-8, a cytokine) is clear.

Our method using a simple ELISA method can drastically reduce thetime and cost, helping speedy product development. Moreover, theq u a n t i t a t i ve m e a s u r e m e n t o f c y to k i n e s e n a b l e s n u m e r i c a lcomparisons in response intensities and time course of the changes
in addition to the determination of presence/absence of response tostimulation, realizing unconventionally efficient examinations.

Skin sensitization assay

01

For the standard 10 compounds of the h-CLAT ( P1 – N10 ), similar results to those in model cells ( LLNA ) can be obtained.
Time and cost can be drastically reduced because our method is free from the time-consuming procedures and confirmation required in the h-CLAT.

Comparison study with the h-CLAT

Our method showed consistent responses to compounds to which inconsistent responses were observed between CD86
and CD54 in the h-CLAT, and our results were similar to those in LLNA.
h-CLATID

Chemical name

LLNA
potencyCD86

iMylc-2
CD54

IL-8Solvent

CAS No.

P1

2,4-Dinitrochlorobenzene

extremePPPDMSO

97-00-7

P2

4-Phenylenediamine

strong

PN

P

EtOH

106-50-3

P3

Nickel sulfate hexahydrate

moderate

PPP

PBS

10101-97-0

P4

2-mercaptobenzothiazole

moderate

Np

P

EtOH

149-30-4

P5R(+) Limonene

weakNpP

EtOH

5989-27-5

P6Imidazolidinyl urea

weak

PPP

PBS

39236-46-9

N7Isopropanol

Non-sensitiser

NNN

PBS

67-63-0

N8Glycerol

Non-sensitiser

NN

N

PBS

56-81-5

N9Lactic acidNon-sensitiser

NN

N

DMSO

50-21-5

N10

4-Aminobenzoic acidNon-sensitiser

NN

N

PBS

150-13-0

11Isoeugenol

Moderate

NNP

97-54-1

121-Bromobutane

Non-sensitiser

PPN

109-65-9

13

Propyl parabenNon-sensitiser

PPN

94-13-3

14

Phthalic anhydride

StrongN

NN

85-44-915

Benzoyl peroxide

StrongNNN

94-36-016

Abietic acid

WeakNNN

514-10-3

17Geraniol

WeakPNN

106-24-1

At present, this assay cannot detect a compound in cases where its metabolites are the cause of sensitivity.

Test methodPyrogen test

02Mylc, which shows similar response to in vivo cells, is superior in sensitivity in sensitizationand stability compared with model cells ( Mono-Mac-6, etc.) used in alternative methods to animal testing.

Comparison study with the Mono-Mac-6

Comparison study in IL-6 production assay, in which a pyrogen ( LPS ) was added. (expressed in OD ) In Mylc-2 ( right figure ),used in this study, the production was detectable at lower concentrations than in Mono-Mac-6 ( left figure ), indicating a higher response.

OD (IL-6)
3.0

OD (IL-6)
3.0
THP-1

2.5

iMylc-2

2.5

Mono-Mac-6

2.0

2.0

1.5

1.5

1.0

1.0

0.5

0.5

0.0

0.0
0.00010.001

0.1101000100000
LPS (pg/mL)

0.001

0.1

101000100000
LPS (pg/mL)

MiCAN Technologies, Inc.

Evaluation and research in acommon laboratory with Kyoto University
Our headquarters and research laboratory are located inKyoto-University Katsura Venture Plaza, where Kyoto University
nurtures the creation of new businesses utilizing new
ideas/technologies and intellectual properties.

0.0001

Kyoto-University Katsura Venture Plaza, 1-36, Goryo-ohara,
Kyoto Nishikyo-ku, Kyoto 615-8245, Japan
Kyoto-University Katsura Venture Plaza

Patent application for non-stimulated dendritic cells for research of viruses (Second product)
Adopted for the Economic Gardening Support Grant supported by Kyoto Industrial Support Organization 21
Certified as Management of Wisdom by the Kyoto Chamber of Commerce and Industry (2018)075-381-3008
http://micantechnologies.com
info@micantechnologies.com