Discovery ofa New Horizon!

-Flow Chemistryto
Human on a Chip-Value

Function

Literature Seminar #2
19/6/2014
Takaya Ukai (M1)

TechnologyConcept

Function

Technology

Value

Function?
3

Innovation

4

Contents
0.#Int'oduction
1.#Flow#Chemist'6

1. Flow Chemistry

2.#Human#on#a#Chip
3.#Discover6#of#a#New#Horizon

5

Goal of This Chapter

First of All…

Hi! This is Akane! I will help
you to follow the discussion.
In this chapter, let’s understandthe “Functions” of Flow
Chemistry. And that’s it!

What is Flow Chemistry?

Assistant:#Akane#Hozuki(a#forensic#scientist)

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Reaction on a Chip

Difference from Flask and Tank

Flow Chemistry= “Reaction on a Chip”“Batch” System“Flow” System

Reagent A
Reagent B

Reagents are mixed together
and react in an ultra narrow tube
(diameter: 100~500 μm)

VSProduct
Aldrich#ChemFiles.#Vol.#9,#No.4,#Microreactor#Technolog6

9Batch System

10Flow System
••
•
•

•

Scale ⇒ Size of the vessel
Each process ⇒ Separated
Heat ⇒ Poorer with Scale↑
Mixing ⇒ Poorer with Scale↑•••Scale ⇒ “Time” of Experiment
Each process ⇒ Connected
Heat ⇒ Almost homogeneous
Mixing ⇒ Almost homogeneous

Heat distribution

Heat distribution

Mixing efficiency

Aldrich#ChemFiles.#Vol.#9,#No.4,#Microreactor#Technolog6

11

Mixing efficiency

Aldrich#ChemFiles.#Vol.#9,#No.4,#Microreactor#Technolog6

12

Advantages of Flow System

And then…

•

Scale Control•

Reaction Control•

Connectable System•

Multistep Synthesis

What can we dowith Flow System?

1314

Scale Control

Reaction Control

Production [g/min] = Flow Rate [mL/min]× Concentration [mol/L]× MW [g/mol]× Yield [%] × 10-3

Controllable Factors:• Temperature (Heat Exchange)• Pressure• Dispersion, Mixing• Reaction time (Retention time)

Scale does not depend on volume but on time

=Aldrich#ChemFiles.#Vol.#9,#No.4,#Microreactor#Technolog6

Highly unstable

15

Kopach,#M.;#Mur'ay,#M.;#Braden,#T.;#Kobierski,#M.;#Williams,#O.#Org.#Process#Res.#Dev.#2009,#13,#152X160.

16

Connectable System

Multistep Synthesis

Connectable Apparatuses:• Column (short pad)
• FLLEX (liquid separation)• ReactIR (reaction monitoring)• HPLC (analysis)• NMR (analysis)
work upreaction monitor&flow rate control

Steven#V.#Ley,#et#al.#Angew.#Chem.#Int.#Ed.#2014,#53,#1

17

Disadvantages of Flow System

Steven#V.#Ley.#The#Chemical#Record.#2012,#12,#378

18

What to Do Next?

Weak Points:• Clogging (viscosity, large particulate)• Gas evolution (shortens retention time)• Pulsating flow
• Corrosion of microreactor

Let’s consider Values and Functionsof Flow Chemistry!

Did you get it?
It’s OK if you grasp the points!

Aldrich#ChemFiles.#Vol.#9,#No.4,#Microreactor#Technolog6

1920

ValuesValue = Advantages (usually)

Flow Chemistry••••

ValueFunction

Scale Control
Reaction Control
Connectable System
Multistep Synthesis

Value for
Organic Chemists

Point: Find out the Value for people
who are engaged in that field

2122

Functions

Functions

Point: Consider what kind of Functions
are essential for the Values

Point: Consider what kind of Functions
are essential for the Values① Inputs are continuously processed and outputted② Construction, fabrication and production

Continuous Systemof micro scale system

Input A

Input B
Processed

Output2324

Functions

Are These Functions Enough?

Point: Consider what kind of Functions
are essential for the Values

Functions:① Inputs are continuously processed and outputted② Construction, fabrication and production③ Flow channels are connectable and recombinable
System Aof micro scale system③ Flow channels are connectable

System Band recombinable

Values:System CSystem D•

Scale Control ⇒ ①•

Reaction Control ⇒ ②, ③•Connectable System ⇒ ③•Multistep Synthesis ⇒ ①, ③

25

Flow Chemistry26?

Function?
Value

Function

Free from Organic Chemistry27?

Innovation?

28

What to Combine with?

Affinity Search

Novelty Conception Approaches:• Forced Creation• Affinity Search• Competence LinkFeasibility

Flow Chemistry•Engineering
Material, Polymer,
Solar Energy,Fuel Chemistry, etc

This time, let’s do “Affinity Search” !
What has high affinity with chemistry?
Can it be a friend with the functions?

•Physics
Quantum Physics,
Thermodynamics,
Nuclear Chemistry, etc

Organic Chemistry•Biology

Biochemistry,
Chemical Biology,
Photosynthesis,Metabolism,
Cellular Biology, etc(Ideal: Indiscriminate trial)

29

Flow’sFunction

30

CellularBiology

2. Human on a Chip
Human on a Chip

31

Goal of This Chapter

Human on a Chip - Destination -Just to confirm how thetechnology is applied tothe other field.
It’s quite easy, isn't it?

33

Human on a Chip - Potential Microsystem which
replicates human bodycompartments utilizing
Microfluid technics

Fully replicate human body functions on a microsystem

34

Replacement of Animal Testing

New assay platform
for drug discovery,
tailor-made medicine,safety test, etc…

35

Animal Testing

Human on a ChipFeeding
Cost

High
(not manufacturable)

Low(manufacturable)

Lengthof Trial

Long

Short

Validity

Insufficient(cross-species)

Sufficient(patient-specific)

A.#Schober,#et#al.#Lab#on#a#Chip.#2013,#13,#3471

36

What Is Necessary?

Prerequisite Basic ResearchesHuman on a Chip

Hmm, the goal is clear though…
Then, why not consider what kinda
basic researches are necessary toachieve that goal? Sorry for saying
nothing special…

Break Down•

Precise replication of each organ•

Vascular system mimetic to connect each organ•

Same standard for tissue engineering

37

Organ on a Chip

=38

First Example: Lung on a Chip
Essential Lung Functions:• Alveolar cellular structure• Contraction when breathing• Inflammatory response

Micro organ composed ofcells with organ-specificfunctions(ex. epithelium, muscle,parenchyma, etc…)=A.#Schober,#et#al.#Lab#on#a#Chip.#2013,#13,#3471

39

Donald#E.#Ingber,#et#al.#Science.#2010,#328,#1662

40

for prolonged periods (>2 weeks) after air was
introduced into the epithelial microchannel and thealveolar cells were maintained at an air-liquidinterface (fig. S1). Addition of air to the upperchannel resulted in increased surfactant production
by the epithelium (Fig. 2B and fig. S1), which
stabilizes the thin liquid layer in vitro as it does inwhole lung in vivo, such that no drying wasobserved. This was also accompanied by an in-Lung on a Chip•

cells not only remained viable but also increased
their surfactant production, enhanced their structural
integrity, and restored normal barrier permeability
in this biomimetic microsystem.
The microfluidic device was then integrated
with computer-controlled vacuum to produce cyclic
stretching of the tissue-tissue interface to mimic
physiological breathing movements (movie S1).
The level of applied strain ranged from 5% toStructure, Contraction•observed in cultured endothelium and in living

microchannel. The presence of these pathogens been difficult in most existing cell stretchingblood vessels in vivo (26, 27). Cyclic stretchingon the apical surface of the alveolar epithelium systems.
causedsomepulsatility
the fluid
flow, but thisIdentification of novel mechanosensitivefor
5 hours
was sufficient
to inactivate
the underunsteadyeffectaswas
negligible
dueof tosmall to nanoparticulates. We also explored
lying
endothelium,
indicatedby capture
cir-theresponses
potential value of this lung-on-a-chip system
culating
neutrophils
theirstretchingtransmigration
into the(seechannelsize andandlow
frequency
for toxicology
applications by investigating the pulthe
alveolarmicrochannel.
reaching theSOMtext).
Thus, this Upon
microdevice
extends
far
alveolar surface, the neutrophils displayed direc- monary response to nanoparticulates delivered tobeyond previously described cell stretching systional movement toward the bacteria, which they the epithelial compartment. Despite the widespread
temsengulfed
by permitting
the applicationof cyclic stretch
use of nanomaterials, much remains to be learned
thenover a periodof a few minutes
and3E
fluidtwoandopposing
layerstheir risks to health and the environment(Fig.
and shearmoviesstress
S8 andtoS9),
the phago-cell about
cytic activity of the neutrophils continued until (31–33). Existing toxicology methods rely on

Lung on a Chip

Inflammatory response*Jargons:
TNF: Inflammatory cytokine
ICAM1: Lymphocyte trafficking signal=

ICAM1 up-regulationb 0 wk=bcutaneous
mplantation

8 weeksthat are commonly used to model the toxic effects
of ultrafine airborne particles (34–36), the underlying endothelium in the microvascular channel
became activated and exhibited high levels ofICAM-1 expression (Fig. 4A). Although physiological breathing movements (10% cyclic strain)
had no effect on ICAM-1 expression on their own,mechanical stretching significantly augmented
endothelial expression of ICAM-1 induced by
the silica nanoparticles (Fig. 4A). Moreover, thiseffect was sufficient to induce endothelial capture&eBM

Neutrophils trapped

Insert eBM

In vivo engineeringof bone marrow

8 wk
Donald#E.#Ingber,#et#al.#Science.#2010,#328,#1662

Fig. 1. Biologically inspired design of a human breathing lung-on-a-chip
microdevice. (A) The microfabricated lung mimic device uses compartmentalized PDMS microchannels to form an alveolar-capillary barrier ona thin, porous, flexible PDMS membrane coated with ECM. The device
recreates physiological breathing movements by applying vacuum to theside chambers and causing mechanical stretching of the PDMS membrane
forming the alveolar-capillary barrier. (B) During inhalation in the living
lung, contraction of the diaphragm causes a reduction in intrapleuralpressure (Pip), leading to distension of the alveoli and physical stretching

Other Organs & Tissues
Paper

www.sciencemag.org

May 2014. Downloaded by Tokyo Daigaku on 13/06/2014 09:02:29.

Gut on a Chip*1dVascular System*2Fig. 3. Reconstitution and direct visualization of complex organ-level responses
irreversibly bonded to form
two sets of three parallel microchannels
involved in pulmonary inflammation and infection in the lung-on-a-chip device.
separated by a 10-mm-thick
PDMSstimulation
membranecontaining
array of(A) Epithelial
with TNF-a(50 ng/ml) an
up-regulates
ICAM-1 expression
through-holes with an effective
ofcontrol10 mm.
200
mm. in the absence(red) on thediameterendothelium;
showsScalelack ofbar,
ICAM-1expressionof TNF-aPDMS
treatment.
Cells were10% strain
0.2 Hz in both cases.
(D) After permanent bonding,
etchant
isstretchedflowedwith
throughtheatside(B) Fluorescently
labeled humanneutrophils(whitechannels
dots) adhere avidly to thechannels. Selective etchingof the membrane
layersin these
activated
endothelium within 1 min after introduction into the vascular channel.
Onlineproduces two large View
sideArticle
chambers
to which vacuum is applied to cause(C) Time-lapse microscopic images showing a captured neutrophil (white arrow)mechanical stretching.
bar, 200mm.
(E) Images
an actualLabScaleon aspreads
Chipthatby firmadhesion
and thenofcrawls
over thelungapical surface of the
on-a-chip microfluidic device
viewedfrom (notabove.
activated
endothelium
visible in this view; direction indicated by yellow

SCIENCE

Bone marrowon a Chip*3

Donald#E.#Ingber,#et#al.#Science.#2010,#328,#1662

41
of the alveolar-capillary interface. (C) Three PDMS layers are aligned andIn Conclusion…

arrows) until it forces itself through the cell-cell boundary within about 2 min

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25 JUNE 2010

1663SCIENCE

www.sciencemag.org

42

after adhesion (times indicated in seconds). During the following 3 to 4 min, theneutrophil transmigrates through the alveolar-capillary barrier by passing
through a pentagonal pore in the PDMS membrane, and then it moves awayfrom the focal plane, causing it to appear blurry in the micrographs. (D) Phasecontrast microscopic images show a neutrophil (arrow) emerging from the apicalsurface of the alveolar epithelium at the end of its transmigration over a period of~3 min; thus, complete passage takes approximately 6 min in total. (E) Timelapse fluorescence microscopic images showing phagocytosis of two GFPexpressing E. coli (green) bacteria on the epithelial surface by a neutrophil (red)that transmigrated from the vascular microchannel to the alveolar compartment.
Scale bar, 50 mm in (A) and (B), 20 mm in (C) to (E).
VOL 3281665

25 JUNE 2010

Did you get it? Though this is justone good successful example, it’s
still helpful to consider “the reason
for success”, I think!

In spite of intensive researches,
this field is still in its infancy
Fig. 4 Photographs of the bioprinted templates (green) enclosed in GelMA hydrogels and the respective microchannels perfused with afluorescent microbead suspension (pink). a) Planar bifurcating bioprinted templates in a GelMA hydrogel construct and (a-i) respective network
after perfusion. b) 3d branching agarose templates embedded in a GelMA hydrogel construct and (b-i) resulting 3d branching network. c) 3d lattice
template embedded in a GelMA hydrogel and (c-i) after perfusion. (scale bars 3 mm, microchannels have 500 μm in diameter).

*1#Geraldine#Hamilton,#Donald#E.#Ingber,#et#al.#Lab#on#a#Chip.#2012,#12,#2165
eBM 8 wk
*2#Ali#Khademhosseini,#et#al.#Lab#on#a#Chip.#2014,#14.#2202
restricting the effective size that constructs can be fabricated.
(Fig. 3D). Upon perfusion *3#Donald#E.#Ingber,#et#al.#Natare#Methods.#2014,#11,#663
of the hydrogel chip with a solution
43
containing rhodamine dye from one inlet (bottom channel),the entire network was rapidly filled (Fig. 3D to D-iii). Suchinterconnection was further demonstrated in a more complex
fluidic network in ESI† movie S4. A potential technical diffi-

We argue that recent strategies our group has developed
enabling directed assembly of microgels, such as using
programmable DNA glue48 or ‘Lego’-like interlocking of
microstructural features in microfabricated microgels, forby clusters of lymphoid cells in the endosteal and perivascular

44

Goal of This Chapter

Discover a frontier far beyond…
3. Discovery ofa New Horizon

In short, understand the whole
thinking process for innovation.
But it’s also crucial to figure out
the limitation of this framework.
Anyway, we are not far off.
Let’s run through toward the end!

46

Affinity Search

Flow Chemistry
Flow’sFunction

Value

CellularBiology

Function

Free from Organic Chemistry47

Human on a Chip

48

Thinking Stream

Is This Framework Applicable?

Find the “interesting” and “novel” technology
Who needs it? How is it useful?
These questions are the keys
probably…
but it’s difficult, I know…hmm

Extract Functions from that technology

Novelty Conception Approaches:• Forced Creation• Affinity Search• Competence LinkFeasibility

Look for the other field which has potential affinity

Combine them and break down to basic studies
49

Who benefits?

50

How Useful?

1. People involved in the Technology

1. People involved in the Technology

Technology

Technology
usually, but not novel

2. People who have a Problem
in their field

Problem

ProblemNovelty

Novelty

3. People who want to do something Novel51

If this process is “Forced Creation”,
it becomes more likely to meet novelty

52

How Useful?
2. People who have a Problem
in their field
Technology

How Useful?
3. People who want to do something Novel

Non-biased search
is important

TechnologyProblem

ProblemNovelty
For them, “Competence Link” isbetter to achieve feasible idea

“Affinity search” is suitable
as a starter for outsiders

Novelty53

Limitation

54

Discover a frontier far beyond…

Creation of a New Research Direction in …
New Research Realm(Blue Ocean)
Existing Research Realm(Red Ocean)It’s difficult to meet all needs.
Complementary process isnecessary, and it’s the next task!

55

Meta Cognition

This is Just an Aside…

Why did we have to investigate thethinking process so intently?
It was abstract and complicated.
…So that I’m hungry.
••…

•Understand what you understand
Understand how you understand
Understand why you understand57

58

Special Thanks

27•Wyss Institute (Harvard University)
for providing Human on a Chip figures•i.school (the University of Tokyo)
for providing hints of the framework•『ぼくは天才ではありません。なぜかというと、自分がどうしてヒットを打てるかを説明できるからです。』

CAPCOMfor dispatching a talented assistant(Akane Hozuki in “Gyakuten-Saiban”)『自分で無意識にやっていることを、もっと意識をしなければならない。』

Thank you very much!
See you next time!

2014 i.school All Rights Reserved.
This material shall be the sole and exclusive property of i.school at The University of Tokyo andshall not be used for any other purpose nor disclosed to any other party without i.school’s writtenconsent.

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