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Mechanobiology Mechanobiology and its Clinical Application and its - - PowerPoint PPT Presentation

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Mechanobiology Mechanobiology and its Clinical Application and its Clinical Application Keiji Keiji Naruse Naruse, M.D. & Ph.D. , M.D. & Ph.D. Professor and Chair Professor and Chair Cardiovascular Physiology, Okayama

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SLIDE 1

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Mechanobiology Mechanobiology and its Clinical Application and its Clinical Application Keiji Keiji Naruse Naruse, M.D. & Ph.D. , M.D. & Ph.D.

Professor and Chair Professor and Chair Cardiovascular Physiology, Okayama Cardiovascular Physiology, Okayama Univ Univ Grad Grad Sch Sch Med Med CSO, STREX Inc. CSO, STREX Inc.

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SLIDE 2

OVERVIEW OVERVIEW

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

MECHANOBIOLOGY MECHANOBIOLOGY MECHANOTRANSDUCTION MECHANOTRANSDUCTION REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

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SLIDE 3

Modified from ”ICURUS” by HENRY MATISSE

HEARING HEARING

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

TOUCH TOUCH JOINTS JOINTS CARDIOVASCULAR CARDIOVASCULAR REPRODUCTION REPRODUCTION

MECHANOBIOLOGY MECHANOBIOLOGY

1xG 1xG

We are continually exposed to mechanical stimuli We are continually exposed to mechanical stimuli

TEETH TEETH

periodontal ligament cells

Bone

slide-4
SLIDE 4

Endothelial cells Endothelial cells

NO, ET1 etc. NO, ET1 etc. Gene expression Gene expression Morphological changes Morphological changes

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Hemodynamic forces Hemodynamic forces

pressure pressure stretch stretch shear stress shear stress STRETCH STRETCH-

  • ACTIVATED

ACTIVATED ION CHANNEL ION CHANNEL

slide-5
SLIDE 5

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Biomedical Engineering Biomedical Engineering MEMS MEMS Biomedical Biomedical

SOFT LITHOGRAPHY SOFT LITHOGRAPHY

  • MEMS(

MEMS(micro electro mechanical systems) )

Mechanobiology Mechanobiology

slide-6
SLIDE 6
  • PDMS

PDMS ( (polydimethylsiloxane polydimethylsiloxane) )

  • nm

nm-

  • mm

mm

  • Merits

Merits

– Transparent

Transparent

– Stable

Stable

– Flexible

Flexible

– High throughput

High throughput

– Inexpensive

Inexpensive

milling milling photolithography photolithography Light, EB Light, EB

photomask photomask regist regist Silicon wafer Silicon wafer

cure cure PDMS PDMS pore pore lift off lift off

Mechanotransduction

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Soft Lithography Soft Lithography

slide-7
SLIDE 7

Stretch chamber Stretch chamber mould mould PDMS PDMS Mechanotransduction

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Chamber Stretch Chamber

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SLIDE 8

Mechanotransduction

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch System Stretch System

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SLIDE 9

1 Hz 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch-

  • induced Morphological change

induced Morphological change

1 Hz 20 %

slide-10
SLIDE 10

0 min 0 min 30 min 30 min 60 min 60 min 120 min 120 min

1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

Stretch Stretch-

  • induced morphological change

induced morphological change

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SLIDE 11

1 Hz, 20 % 60 min 1 Hz, 20 % 60 min Ca Ca2+

2+ free

free Gd Gd3+

3+

Gd Gd3+

3+

Ca Ca2+

2+ free

free

1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

0 min 0 min 30 min 30 min 60 min 60 min 120 min 120 min

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

30 60 90 120 150 180 5 10 15 20

%

Stretch Stretch-

  • induced morphological change

induced morphological change

Stretch Stretch-

  • activated channel dependent

activated channel dependent

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SLIDE 12

Plasma membrane Plasma membrane SA channel SA channel Ca Ca2+

2+influx

influx

Fura2 Fura2

Fluorsecence Fluorsecence Microscopy Microscopy

Gd blocks SA channel 1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch-

  • induced Ca response

induced Ca response

slide-13
SLIDE 13

1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch-

  • induced Ca response

induced Ca response

slide-14
SLIDE 14

(10%) (5%) (15%) (20%) (25%)

Br J Pharmacol, 2002

200 400 600 800 Time (sec) 0.4 0.6 0.8 1.0 1.2 1.4 1.6 Ratio (340/380)

Stretch for 3 sec

1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch-

  • induced Ca response

induced Ca response

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SLIDE 15

100 200 300 400 500 600 0.4 Time (sec)

Gd3+

(10%) (5%) (15%) (20%) (25%) (10%) (5%) (15%) (20%) (25%)

Br J Pharmacol, 2002

0.4 0.6 0.8 1.0 1.2 1.4 Ratio (340/380)

Stretch for 3 sec

1 Hz, 20 %

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch-

  • induced Ca response

induced Ca response

Stretch Stretch-

  • activated channel dependent

activated channel dependent

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SLIDE 16
  • MscL

MscL, , MscS MscS (Non (Non-selective) selective)

  • SAKCA (K

SAKCA (K + selective) selective)

  • Mid

Mid-

  • 1 (

1 (Non Non-

  • selective)

selective)

  • TREK/TRAAK

TREK/TRAAK ( (K+ selective) selective)

  • TRPs (

TRPs (cation cation selective) selective)

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

slide-17
SLIDE 17

Science, 285:882 Science, 285:882-885 1999 885 1999

Molecular Identification of a Eukaryotic, Stretch Molecular Identification of a Eukaryotic, Stretch-

  • Activated Nonselective

Activated Nonselective Cation Cation Channel Channel ：Mid1 Mid1

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

Mid1 Mid1

slide-18
SLIDE 18

MDALIIPVTMEVPCDSRGQRMWWAFLASSMVTFFGGLFIILLWRTLKYLW 50 MDALIIPVTMEVPCDSRGQRMWWAFLASSMVTFFGGLFIILLWRTLKYLW 50 TVCCHCGVKNKEAQKINGGGDTQADGACKPTDEKEENVAAEVGWMTSVKD 100 TVCCHCGVKNKEAQKINGGGDTQADGACKPTDEKEENVAAEVGWMTSVKD 100 WAGVMISAQTLTGRVLVVLVFALSIGALVIYFIDSSNPIESCQNFYKDFT 150 WAGVMISAQTLTGRVLVVLVFALSIGALVIYFIDSSNPIESCQNFYKDFT 150 LQIDMAFNVFFLLYFGLRFIAANDKLWFWLEVNSVVDFFTVPPVFVSVYL 200 LQIDMAFNVFFLLYFGLRFIAANDKLWFWLEVNSVVDFFTVPPVFVSVYL 200 NRSWLGLRFLRALRLIQFSEILQFLNILKTSNSIKLVNLCSIFISTWLTA 250 NRSWLGLRFLRALRLIQFSEILQFLNILKTSNSIKLVNLCSIFISTWLTA 250 AGFIHLVENSGDPWENFQNNQQLTYWECVYLLMVTMSTVGYGDVYAKTTL 300 AGFIHLVENSGDPWENFQNNQQLTYWECVYLLMVTMSTVGYGDVYAKTTL 300 GRLFMVFFILGGLAMFASYVPEIIELIGNRKKYGGSYSAVSGRKHIVVCG 350 GRLFMVFFILGGLAMFASYVPEIIELIGNRKKYGGSYSAVSGRKHIVVCG 350 HITLESVSNFLKDFLHKDRDDVNVEIVFLHNISPNPELEALFKRHFTQVE 400 HITLESVSNFLKDFLHKDRDDVNVEIVFLHNISPNPELEALFKRHFTQVE 400 FYQGSVLNPHDLARVKIESADACLILANKYCADPDAEDASNIMRVISIKN 450 FYQGSVLNPHDLARVKIESADACLILANKYCADPDAEDASNIMRVISIKN 450 YHPKIRIITQMLQYHNKAHLLNIPSWNWKEGDDAICLAELKLGFIAQSCL 500 YHPKIRIITQMLQYHNKAHLLNIPSWNWKEGDDAICLAELKLGFIAQSCL 500 APGLSTMLANLFSMRSFIKIEEDTWQKYYLEGVANEMYTEYLSSAFVGLS 550 APGLSTMLANLFSMRSFIKIEEDTWQKYYLEGVANEMYTEYLSSAFVGLS 550 FPAVCELVFAKLKLLMIAIEYKSEKRESSILINPGNHVKIQEGTLGFFIA 600 FPAVCELVFAKLKLLMIAIEYKSEKRESSILINPGNHVKIQEGTLGFFIA 600 SDAKEVKRAFFYCKACHDDITDPKRIKKCGCKR SDAKEVKRAFFYCKACHDDITDPKRIKKCGCKRPKMSIYKRMKLACCFDC PKMSIYKRMKLACCFDC 650 650 GRSERDCSCMSGSVHSNMDTLERAFPLSSVSVNDCSTSLRAF GRSERDCSCMSGSVHSNMDTLERAFPLSSVSVNDCSTSLRAFEDEQPSTL 700 EDEQPSTL 700 SPKKKQRNGGMRNSPNSSPKLMRHDPLLIPGNEQIDNMDANVKKYDSTGM 750 SPKKKQRNGGMRNSPNSSPKLMRHDPLLIPGNEQIDNMDANVKKYDSTGM 750 FHWCPAKDIEKVILTRSEAAMTVLSGHVVVCIFGDVKSALIGLRNLVMPL 800 FHWCPAKDIEKVILTRSEAAMTVLSGHVVVCIFGDVKSALIGLRNLVMPL 800 RASNFHYHELKHIVFVGSLEYLRREWETLHNFPKVSILPGTPLSRADLRA 850 RASNFHYHELKHIVFVGSLEYLRREWETLHNFPKVSILPGTPLSRADLRA 850 VNINLCDMCVILSANQNNIDDASLQDKECILASLNIKSMQFDDSIGVLQA 900 VNINLCDMCVILSANQNNIDDASLQDKECILASLNIKSMQFDDSIGVLQA 900 NSQGFTPPGMDRSSPDNSPVHGLLRQPSITTGANIPIITELVNDSNVQFL 950 NSQGFTPPGMDRSSPDNSPVHGLLRQPSITTGANIPIITELVNDSNVQFL 950 DQDDDDDPDTELYLTQPFACGTAFAVSVLDSLMSATYFNDNILTLIRTLV 1000 DQDDDDDPDTELYLTQPFACGTAFAVSVLDSLMSATYFNDNILTLIRTLV 1000 TGGATPELEALIAEENALRGGYSTPQTLANRDRCRVAQLALYDGPFADLG 1050 TGGATPELEALIAEENALRGGYSTPQTLANRDRCRVAQLALYDGPFADLG 1050 DGGCYGDLFCKALKTYNMFCFGIYRLRDAHLSTPSQCTKRYVITNPPYEF 1100 DGGCYGDLFCKALKTYNMFCFGIYRLRDAHLSTPSQCTKRYVITNPPYEF 1100 ELVPTDLIFCLMQFDHNAGQSRASLSHSSHSSYSSSKKSSSVHSIPSTAN 1150 ELVPTDLIFCLMQFDHNAGQSRASLSHSSHSSYSSSKKSSSVHSIPSTAN 1150 RPNRTKTRDSREKQKYVQEDRL 1172 RPNRTKTRDSREKQKYVQEDRL 1172

SAKCA

STREX: STREX: St Stress Axis ress Axis Regurated egurated EX EXon

  • n

`

STREX

S0S1S2S3S4S5 S6

COOH NH2

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

10 20 30 40 5 10 15 20 25 30 pressure(-m mHg) Po (%) 50 `

STREX

S0 S1 S2 S3 S4 S5S6

`

S0 S1 S2 S3 S4 S5 S6

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SLIDE 19

TRPM TRPP TRPML TRPC TRPV TRPA TRPN

Mechanosensitive channel

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

Transient Receptor Potential channels

・ Drosophila Retina ・ non-selective cation channel ・ 6 putative transmembrane helices ・ ankyrin repeats (N-terminal) ・ variety of sensory system ・ TRPV1→Thermosensor ・ Mechanosensor? ・ TRPV2→SA channel?

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SLIDE 20

COS7（Control）

10% 20% 40%

10 20 40 COS7（hTRPV2-His） 10%

20% 40%

10 20 40

Transient Rceptor Potential channels

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

COS7（hTRPV2-His）

SA channel SA channel

TRPV2 TRPV2

slide-21
SLIDE 21

1

100000 200000 0.3 0.4 0.5 0.6 0.7 0.8

Ratio (340nm/380nm) Time (msec)

4 5 6 2 3

1 2 3 4 5 6

ratio 380 nm TRPV2-siRNA

Effect of TRPV2-siRNA on stretch-induced [Ca2+]i response

HUVEC

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

TRPV2 TRPV2

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SLIDE 22

心筋症・心不全 におけるTRPV2の役割

NE NEW Y YORK, M , Manhatta ttan

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

TRPV2 TG, KO, KI mouse TRPV2 TG, KO, KI mouse

Inducible, Tissue specific, Human KI Double KO Wild V2 TG

slide-23
SLIDE 23

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel

TRPV2 TRPV2

Ito et al., ARCMB, 2008

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SLIDE 24

“Nano-crack”

Fabrication of reconfigurable protein matrices by cracking

Nature Materials 4（5）：403‐406，2005

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

MECHANOTRANSDUCTION MECHANOTRANSDUCTION

slide-25
SLIDE 25

Conventional cantilever

Fast Scanning cantilever

10μ 10μｍ ｍ

200μ 200μｍ ｍ

Olympus

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Fast Scanning Atomic Force Microscopy Fast Scanning Atomic Force Microscopy

plasmid

(800 x 600 nm)

Fibronectin

(７４0 x 560 nm)

1 nm 10 nm

MscL / WT MscL / G22N

Open mutant

5 nm

slide-26
SLIDE 26

Mold & cure

mold cure

Ink Print

PDMS stretch-chamber Fibronectin PDMS Fibronectin

(A) (B) (C) (D) (E) (F) (G) (H)

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Microcontact Printing

Stretch Stretch-

  • induced

induced Responses Responses in Patterned in Patterned HUVEC HUVEC

slide-27
SLIDE 27

Before stretch After stretch (15 min) After stretch (60 min) Anti- PY Phalloidine

20 um

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Microcontact Printing on Stretch chamber

Stretch Stretch-

  • induced

induced Responses Responses in Patterned in Patterned HUVEC HUVEC

0.1 0.2 0.3 0.4 0.5 0.6 0.7

Ratio (A340/A380)

stretch

35μm×35μm 25μm×25μm 20μm×20μm 10μm×10μm

1min

Z X-Y

Fura 2

Tyrosine phosphorylation Stretch-induced [Ca2+]i

J Biotech 2008

slide-28
SLIDE 28

Coverslip

Cells in microchannel

W=200μm H=50μm

①High shear ②Laminar flow

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

MECHANOTRANSDUCTION MECHANOTRANSDUCTION

slide-29
SLIDE 29

CONTROL SHEAR-STRESS

Nuclear Factor kB

Cell response in microchannel

0.4 0.5 0.6 0.7 0.8 0.9 1 500 600 700 800 900 1000

time (sec) r at io ( F 3 4 0 /F 3 8 0 )

0.25μM ATP 9 dyn/ cm2 1 8 dyn/ cm2

[Ca2+]i in response to shear stress

0.4 0.5 0.6 0.7 0.8 0.9 1 500 600 700 800 900 1000

time (sec) r at io ( F 3 4 0 /F 3 8 0 )

0.25μM ATP 9 dyn/ cm2 1 8 dyn/ cm2

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

MECHANOTRANSDUCTION MECHANOTRANSDUCTION

slide-30
SLIDE 30

Principle of laminar flow

～500μm in width ～ 50μm in hight W=500μm H=50μm

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

MECHANOTRANSDUCTION MECHANOTRANSDUCTION

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SLIDE 31

MECHANOTRANSDUCTION MECHANOTRANSDUCTION

W=200μm H=50μm

Principle of laminar flow

PARTCEL (pattented) “Subcellular positioning of small molecules” Nature, 2001

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

A B C

slide-32
SLIDE 32

Human uman Assisted ssisted Reproductive eproductive Therapy herapy

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

INFERTILITY INFERTILITY

Mechanobiology Mechanobiology in Reproductive Medicine in Reproductive Medicine

slide-33
SLIDE 33
  • Decreasing in Population

Decreasing in Population

  • Death(

Death(1,077,000

1,077,000)>Birth(

)>Birth(1,067,000

1,067,000)

  • Decreasing in Birthrate

Decreasing in Birthrate

  • Total fertility rate : ~1.3

Total fertility rate : ~1.3

  • Various cause

Various cause

  • Infertility

Infertility

  • One in 5

One in 5-

  • 10 couples

10 couples

Male factor=Female factor Male factor=Female factor

  • 2-
  • 3 % : IVF

3 % : IVF

平成18年6月2日日経新聞

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

INFERTILITY in JAPAN INFERTILITY in JAPAN

Mechanobiology Mechanobiology in Reproductive Medicine in Reproductive Medicine

slide-34
SLIDE 34

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

INFERTILIGY TREATMENT INFERTILIGY TREATMENT

  • Timing treatment (Ovulation)

Timing treatment (Ovulation)

  • AIH (Artificial Insemination by Husband)

AIH (Artificial Insemination by Husband)

  • IVF (In Vitro Fertilization)

IVF (In Vitro Fertilization)

– – IVF IVF‐conventional conventional – – IVF IVF‐ ‐ICSI ICSI（Intracytoplasmic Intracytoplasmic sperm injection sperm injection）

Sperm Artificial Insemination Conventional IVF ICSI

slide-35
SLIDE 35

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Conventional Conventional Human uman Assisted ssisted Reproductive eproductive Treatment (HART) reatment (HART)

Density gradient Swim‐up

centrifuge 400×g, 20min

Sperm preparation Sperm preparation

Culture dish Mimeral oil Culture medium Embryo

STATIC CULTURE Egg culture Egg culture

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE Non Non-

  • physiological

physiological

slide-36
SLIDE 36
  • 1. Microfluidic Sperm Sorter System
  • 2. Mechanically Active Embryo Culture System

STREX www.strex.jp www.strex.jp

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-37
SLIDE 37

STREX www.strex.jp www.strex.jp

W=500μm, H=50μm

Sperm Sorter /microfluidics

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-38
SLIDE 38

Takayama S, Analytical Chemistry, 75:1671-1675, 2003 STREX www.strex.co.jp www.strex.co.jp Initial Initial Sorted Sorted

20 20 40 40 60 60 80 80 100 100

Sperm Motility (%) Sperm Motility (%)

n=10 n=10 n=10 n=10

a b

a,b; P<0.05 a,b; P<0.05

Sperm Sorter /microfluidics

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-39
SLIDE 39

400 x ｇ

Conventional protocol

Motile sperm

SWIM UP !! SWIM UP !!

MFSS & Dilution

+

semen medium Dilution > X 3 MFSS

No centrifugation

＜岡山大学医学部＞ ＜名古屋大学医学部＞

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

No Centrifugation No Centrifugation

slide-40
SLIDE 40

conv MFSS separation min ０ ３０ ６０ ９０ １２０ semen prep centrifugation density grad Swim up ＜名古屋大学医学部＞

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

Short processing time Short processing time

slide-41
SLIDE 41

10 20 30 40 50 60 70

%

59.6% 59.6% 59/99 59/99 46.8% 46.8% 51/109 51/109 Sperm Sorter Sperm Sorter conventional conventional

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

Fertilization rate ↑ Fertilization rate ↑

  • IRB: Passed 13/5/2005
  • 40 couples with

informed consent

  • > 4 eggs ⇒ ICSI
  • Embryo transplantation
slide-42
SLIDE 42

＜プラスチック製＞

0.45 mm 0.08 mm 1 JPY

Quartz Plastic

0.49 mm 0.18 mm

a b c d

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-43
SLIDE 43

http://www2.edu.ipa.go.jp/gz/

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-44
SLIDE 44

http://www2.edu.ipa.go.jp/gz/

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

RAT FALLOPIAN TUBE RAT FALLOPIAN TUBE Mouse eggs in fallopian tube Mouse eggs in fallopian tube

slide-45
SLIDE 45

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Shear Stress Compression Stretch Turbulence

Friction

Mechanical stresses applied to embryo in oviduct Mechanical stresses applied to embryo in oviduct

REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE

slide-46
SLIDE 46

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

TECS TECS

Tilting Embryo Culture System (TECS) Tilting Embryo Culture System (TECS)

Shear stress (dynes/cm2) Velocity (mm/sec) 0.003

0.0018

*p=0.035

Blastocysts/total % Control 63/136 46 TECS 77/130 59 Tilting culture unit Control unit

STREX Inc. 20deg tilting for 10 min, 4-6 mouse eggs/drop

Roll

slide-47
SLIDE 47
  • Nagoya Univ

– M. Sokabe – T. Yamada – X. Sai – K. Wang – H. Inoh – H. Takeda

  • Nagoya Univ

Hospital

– H. Ando – D. Shibata

  • Harvard Univ

– D. Ingber – G.Whitesides

  • Michigan Univ

– S. Takayama – G. Smith

  • ICORP/JST

– X. Tang – F. Lopez

  • OKAYAMA Univ

– S. Mohri – Y. Katanosaka – G. Iribe – A. Yamada – H. Kishio – H. Bao – T. Suemori – T. Komatsu – M. Kajiya

Acknowledgement

Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine