Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

Alignment of Myoblast Cultured on Micro Striped Ridge After
Centrifuge Stimulation: Before and After Division
Yuji ENDO, Shigehiro HASHIMOTO, Yoshisato KAGEYAMA, Yu MORITA
Biomedical Engineering, Department of Mechanical Engineering, Kogakuin University,
Kogakuin University, Tokyo, 163-8677, Japan
http://www.mech.kogakuin.ac.jp/labs/bio/
Kiyoshi YOSHINAKA
Health and Medical Research Institute, National Institute of Advanced Industrial Science & Technology
Tsukuba, 305-8564, Japan
ABSTRACT

differentiation and growth of C2C12 were delayed with electricpulses in the previous study in vitro [7]. The floating cells of
C2C12 were tried to be sorted by the micro groove on the wallof the flow channel [8].

The effect of hysteresis of the tangential force field at thesurface of the scaffold on the alignment of myoblast has beenstudied in vitro. The striped pattern (0.7 μm height, 3 μm
width, and 3 μm interval) were made on the surface of thescaffold plate to control the orientation of each cell. Variation
was made on the angle between the longitudinal direction of theridge and the direction of the tangential force: 0 degree, 45
degrees, and 90 degrees. C2C12 (mouse myoblast cell line)
was used in the experiment. To apply the tangential force field(< 100 G) to the cells, the scaffold plate was set in the tube in aconventional centrifugal machine placed in an incubator.
After the centrifugation for 5 hours, the behavior of each cell at
the time-lapse microscope images was traced for 24 hours toanalyze the angle between the longitudinal axis of the cell andthe direction of the past centrifugation. The experimental
results show that the tendency of cells to align the longitudinal
direction of the striped pattern is strengthened by the hysteresisof the exposure to the tangential force field. The tendency isstrengthened at the division of each cell.

These behaviors might depend on the history of each cell.
While the strong stimulation above the threshold damages thecell, the stimulation below the threshold remains in the cell as amemory for the response in the next step. The memory might
be reset after division of the cell. To trace the hysteresis effectof the mechanical stimulation on the single cell, the time-lapse
images are effective. In the previous study, the longitudinal
axis of C2C12 (mouse myoblast cell line) tends to align to thedirection of the excess gravity.
The axis tilts to theperpendicular direction, on the other hand, after stopping of theexcess gravity [9]. A single cell migrates at random on thescaffold. The cell tends to align to the longitudinal directionof the micro ridge line [10].
In the present study, centrifugal force has been used to apply thetangential force field on the surface of the scaffold in vitro.
The hysteresis effect of the force field on the single cellattached on the scaffold has been investigated, after stopping of
the stimulation of the excess gravity. To examine the effect ofthe direction of the force field on the cell at the specificalignment, the orientation of the cell was controlled by the
striped pattern on the surface of the scaffold. The behavior ofeach cell has been traced before and after division.

Keywords: Biomedical Engineering, Cell Culture, Excess
Gravity, Orientation and C2C12.

1. INTRODUCTION
The recent cell culture technique enables observation of the
behavior of each cell in vitro. A biological cell adheres on thescaffold and shows several active behaviors: migration,
deformation, and proliferation. The cell is deformed by theforce, because of its compliance. The cell deforms, on theother hand, to minimize the intra force of itself. The cell is
moved by the force. The cell, on the other hand, moves byitself. C2C12 is cell line of mouse myoblast. In the previous
study, both the migration and the deformation of C2C12 wererestricted by the wall shear stress higher than 3 Pa in flow invitro [1]. Deformability of C2C12 was observed by the slit in
the previous study in vitro [2, 3]. C2C12 extended along thelines of micro ridges on the scaffold surface [4]. Vibration
decelerated adhesion of C2C12 on the micro ridges [5].
C2C12 migrated regardless of the direction of the vibrating
micro ridges. The orientation of C2C12 was tried to bemeasured by the electric impedance in vitro [6]. Both

2. METHODS
Micro-pattern on Scaffold Plate
The micro striped pattern (Fig. 1) has been made in three partial
rectangular areas of 0.4 mm × 1.6 mm on the PDMS
(polydimethylsiloxane)
plate
of
the
scaffoldbyphotolithography technique. The height (H), the width (W),and the interval (I) of the quadrangular ridges are 0.7 μm, 3 μm,
and 3 μm, respectively. Each area has its own specificdirection of the striped pattern. Namely, variation has been
made on the angle (α) between the longitudinal direction of theridge and the direction of centrifuge: 0 degree (parallel), 45
degree, and 90 degree (perpendicular). Three partial area wasmade on the same surface of the scaffold plate in parallel
position, so that the behavior of cells on each area can be7

Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

apply the excess gravity, the plate, on which cells were cultured,
was set in the medium in the tube. Several tubes with theglass plate were set in the rotor of the centrifugation to cultivate
cells, simultaneously. To keep the content of carbon dioxideof 5 % at 310 K, the cells were cultured in an incubator throughthe entire experimental term including the term of exposure to
the hyper-gravitational field. Namely, the centrifugal machine
was placed in the incubator during the centrifugation.

compared simultaneously (Fig. 2). The pattern of each area
was also used as a marker to trace each cell.
Mold for Micro Pattern
The borosilicate glass (Tempax) disk was used for the base of
the mold through micromachining process. The positive
photoresist material of OFPR-800 (Tokyo Ohka Kogyo Co., Ltd,Tokyo, Japan) was coated on the glass with the spin coater (at3000 rpm for 20 s). The micro pattern was drawn on the moldwith a laser drawing system (DDB-201K-KH, Neoark
Corporation, Hachioji, Japan). The photoresist was developedwith tetra-methyl-ammonium hydroxide (NMD-3, Tokyo Ohka
Kogyo Co., Ltd., Kawasaki, Japan) for two minutes. The glass
plate with the photoresist material was etched with the plasma
gas using RIE-10NR (Samco International, Kyoto, Japan).

In the control group, the cells were cultured without centrifuge
on the plate, which was placed in the polystyrene dish. In thecontrol group, no tangential force field was applied, and only
the gravitational force (1 G) normal to the scaffold surface plate
was applied.
Image Analysis
After stimulation of the excess gravity, cells on the glass plate
were moved from the centrifugal tube to the culture dish.
Cells were subsequently cultured in the dish placed in the
incubator combined with an inverted phase-contrast microscope(LCV110-SK, Olympus Co., Ltd., Tokyo). The behavior ofeach cell was analyzed on the time-lapse images captured every
ten minutes for 24 hours after stimulation of the excess gravity.
Cells, which made division, were traced: before and after
division. “Image J” was applied to analyze the behavior ofeach cell. On the microscopic image, the outline of each cell
was traced, and the contour of each cell was approximated toellipsoid (Fig. 1). On the ellipsoid, the length of the major
axis (a), and the minor axis (b) were measured. The ratio ofaxes is calculated as the shape index (P) by Eq. (3).

After the mold of the glass disk was enclosed with a peripheral
wall of polyimide tape, PDMS (Sylgard 184 Silicone Elastomer
Base, Dow Corning Corp., MI, USA) was poured with thecuring agent (Sylgard 184 Silicone Elastomer Curing Agent,
Dow Corning Corp., MI, USA) on the mold. The volume ratioof PDMS to curing agent is ten to one. The baked plate of
PDMS (1 mm thickness) was exfoliated from the mold of theslide glass plate. The PDMS plate was cut to make a plate of
15 mm × 10 mm × 1 mm, and stacked on the glass plate of 50
mm × 13 mm × 1 mm. The surface of the PDMS plate was
exposed to the oxygen gas (0.1 Pa, 30 cm3/min) in the reactive
ion etching system (FA-1: oxygen plasma ashing, 50 W) for
thirty seconds just before the cell culture.
Tangential Force Field
The tangential force field was applied to the culture surface
with the centrifugal force. The culture plate is inserted into thetube, which is contained in the rotor. The angle between theradial direction of the rotation of the rotor and the axial
direction of the tube in the rotor is 60 degree. The glass plate
was set so that the culture surface is parallel to the radial
direction of the rotation. The centrifugal force is applied
parallel to the culture surface. The centrifugal force (Fc) is
calculated by Eq. (1).
Fc = m r ω 2

P=1 㸫 b/a(3)

At the circle (a = b), P = 0. As the ellipsoid is elongated (a >>b), P approaches to 1. The acute angle (0 < θ < 90 degree)
between the direction of the centrifuge and the direction of the
major axis of each cell was measured at the microscopic image.
When the major axis is parallel to the direction of centrifuge, θ= 0. When the major axis is perpendicular to the direction ofcentrifuge, θ = 90 degree.

(1)
3. RESULTS

In Eq. (1), m is mass, r is radius of the rotation, and ω is angularvelocity. In the gravitational field, gravitational force (Fg) is
calculated by Eq. (2), where g is gravitational acceleration.
Fg = m g

By the time-lapse images, continuous activity of each cell was
confirmed: migration, deformation, and division (Fig. 2).
Each cell repetitively extends pseudopods. The centrifuge
direction was easily found by the stripe patterns of 0 degree.
The behavior of each cell was easily traced with the help of the
stripe patterns as markers. Each cell shows active behavior on
micro striped ridges: migration, deformation, and division.

(2)

In the present study, the centrifugal acceleration lower than 100
G (1 G is equal to the gravitational acceleration) ((Fc / Fg) =100) is applied with the centrifugal machine (CN-1040,
Matsuuraseisakusyo. Ltd, Tokyo, Japan).

20 µm

Cell Culture
C2C12 (mouse myoblast cell line originated with cross-striatedmuscle of C3H mouse) was used in the test. C2C12 of thepassage four was cultured in D-MEM (Dulbecco’s Modified
Eagle’s Medium) containing 10% decomplemented FBS (fetal
bovine serum) and 1% penicillin/ streptomycin. The cells
were seeded on the glass plate at the density of 3000 cells/cm2.
After the cells were cultured for 12 hours in the resting state,excess gravitational stimulation was applied for five hours.
The gravity of 50 G (r = 0.045 m, group X), and the gravity of
100 G (r = 0.09 m, group Y) were applied at ω = 104 rad/s. To

a

Cell
b

θ
Centrifuge
Fig. 1: On microscopic image, outline of each cell was traced,and contour of each cell was approximated to ellipsoid: on
stripe ridge lines (α = 90 degree).

8Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

Figs. 3–5 show tracings of the angle (θ) of each cell. Each cell
divides at the time zero. Each tracing of two tracings after
division show tracing of each divided cell. Fig. 3 shows theangle of cell without centrifugal stimulation. Without thetangential force stimulation, the angle (θ) of each cell tends toalign to the direction of striped ridges (Fig. 3). The tendency
to align parallel to the tangential force field does not affect thealignment of each cell one hour after stimulation (Figs. 4 & 5).
Each cell tends to align to the direction of the striped ridges,
except just after division. On the striped ridges parallel to thetangential centrifugal force field of 100 G, each cell remarkablyaligns to the direction of the striped ridges after the division(Fig. 5a). Cells often make opposite alignment each other justafter the division (Figs. 3b & 4b).

Angle (θ)( ) [degree]Time [min]Fig. 3b: Angle (θ) of C2C12: control, α = 45 degree.
Angle (θ) [degree]

Figs. 6–8 show tracings of the shape index (P) of each cell.
The shape index (P) decreases immediately after division,
which corresponds to approaching to the spherical shape. On
the striped ridges parallel to the tangential centrifugal force
field of 100 G, on the other hand, the shape index increases
occasionally (Fig. 8a).
Data are rearranged as the relationships between the shapeindex (P) and the angle (θ) of each cell after 100 G in Fig. 9.
The regression line shows the tendency that the shape index (P)decreases with the angle (θ) on the striped ridges of 0 degree (α)(Fig. 9a). The tendency corresponds to the elongation of thecell to the longitudinal direction of the ridge. The correlation
coefficient (r2) is higher before the cell division. The tendency
is not clear after the cell division: small value of the correlation
coefficient. The decrease of the shape index (P) with the angle(θ) is not clear in Fig. 9b. That corresponds to the unclear
tendency of the elongation of the cell to the longitudinal
direction of the ridge (α = 90 degrees).

90 degTime [min]Fig. 3c: Angle (θ) of C2C12: control, α = 90 degree.
Angle (θ) [degree]Time [min]Centrifugal force

Fig. 4a: Angle (θ) of C2C12: 50 G, α = 0 degree.
Angle (θ) [degree]

45 deg0 deg

0.2 mmFig. 2: Cells after stopping centrifuge: α = 90 degree (upper),45 degree (middle), and 0 degree (lower): dotted line shows
direction of ridges.

Time [min]Angle (θ) [degree]

Fig. 4b: Angle (θ) of C2C12: 50 G, α = 45 degree.
Angle (θ) [degree]Time [min]Fig. 3a: Angle (θ) of C2C12: control, α = 0 degree.
Time [min]Fig. 4c: Angle (θ) of C2C12: 50 G, α = 90 degree.

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Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

Shape index (P)Angle (θ) [degree]Time [min]Fig. 6c: Shape index (P) of C2C12: control, α = 90 degree.

Time [min]Fig. 5a: Angle (θ) of C2C12: 100 G, α = 0 degree.

Shape index (P)Angle (θ) [degree]Time [min]Fig. 7a: Shape index (P) of C2C12: 50 G, α = 0 degree.

Time [min]Fig. 5b: Angle (θ) of C2C12: 100 G, α = 45 degree.

Shape index (P)Angle (θ) [degree]Time [min]Time [min]Fig. 5c: Angle (θ) of C2C12: 100 G, α = 90 degree.

Fig. 7b: Shape index (P) of C2C12: 50 G, α = 45 degree.
Shape index (P)Shape index (P)Time [min]Time [min]Fig. 6a: Shape index (P) of C2C12: control, α = 0 degree.

Fig. 7c: Shape index (P) of C2C12: 50 G, α = 90 degree.

Shape index (P)Shape index (P)Time [min]Time [min]Fig. 6b: Shape index (P) of C2C12: control, α = 45 degree.

Fig. 8a: Shape index (P) of C2C12: 100 G, α = 0 degree.

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Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

4. DISCUSSION

Shape index (P)The hysteresis of centrifugation might make each cell increase
the area contact with the surface of the scaffold. Each cellmight adapt to the tangential force field by maximization of thecontact area between the cell and the scaffold.
To control the alignment of the single cell on the scaffold before
stimulation, the micro stripe pattern has been used in the present
study. The height of the ridge is designed as 0.7 μm, which isslightly higher than threshold to make orientation of cells [10].
The average diameter of the floating cell in the medium is 20
μm. The low ridge and the narrow width were selected so as
not to disturb cell migration to every direction.
Bothdeformation and migration of C2C12 are at random over the
striped pattern in the present study.

Time [min]Fig. 8b: Shape index (P) of C2C12: 100 G, α = 45 degree.
Shape index (P)C2C12, on the other hand, tended to elongate and align along
the striped pattern even in the case without the tangential force
stimulation in the previous study [9]. The tilting tendency of
C2C12 was enhanced by the tangential force of 100 G. C2C12
tends to be elongated during the tangential force stimulation.
The mild tendency was observed on MC3T3-E1 in the previous
study [11].
The behavior of each cell can be easily traced with the aid of
the micro striped pattern on the scaffold as the marker. The
tracking ability both to the tangential force and to surface
morphology was higher in C2C12 than in MC3T3-E1 in theprevious study. The tracking ability of 3T3-L1 is much lower
than that of MC3T3-E1 [11]. On the striped pattern parallel tothe tangential force (0 degree), every type of cell is elongatedalong the striped pattern. The tracking ability on anotherdirection of striped pattern depends on the cell types.

Time [min]Fig. 8c: Shape index (P) of C2C12: 100 G, α = 90 degree.
Shape index (P)In the tangential force field, each cell tends to make stronger
contact to the scaffold, which makes orientation of cells along
the micro striped patterns parallel to the tangential force. In
the previous study, C2C12 was most frequently elongated on
the striped pattern of 45 degree between the tangential force
direction and the longitudinal direction of the stripe pattern [9].
Angle (θ) [degree]

C2C12 differentiation might be accelerated by the mechanicalstimulation in the tangential force field by the centrifugal field.
The myotube might align to the direction perpendicular to thecentrifugal field. Hypergravity affects myoblast [11, 12].
Hypergravity affects tissue formation [13, 14]. The effect wasstudied with design of scaffold [15].
The effect ofhypergravity depends on cell types [16]. The effect of
mechanical stress affects molecular response [17–20]. The
acceleration technique for orientation, proliferation anddifferentiation of cells has been studied to make tissue in vivo orin vitro. Control methodology for orientation, proliferationand differentiation of cells would be applied to the regenerative
tissue technology.

Fig. 9a: Shape index (P) vs. angle (θ) of C2C12: 100 G, α = 0
degree (Fig. 5a, 8a): before (asterisk, r2 = 0.45), after (plus, r2 =0.23; cross, r2 = 0.08) division.
Shape index (P)5. CONCLUSION
Angle (θ) [degree]
Fig. 9b: Shape index (P) vs. angle (θ) of C2C12: 100 G, α = 90
degree (Fig. 5c, 8c): before (asterisk, r2 = 0.09), after (plus, r2 =0.25; square, r2 = 0.04) division.

The effect of hysteresis of the tangential force field at thesurface of the scaffold on the alignment of myoblast has beenstudied in vitro. The striped pattern (0.7 μm height, 3 μm
width, and 3 μm interval) were made on the surface of thescaffold plate to control the orientation of each cell. To apply
the tangential force field (< 100 G) to C2C12 (mouse myoblast11

Proceedings of The 24th World Multi-Conference on Systemics, Cybernetics and Informatics (WMSCI 2020)

cell line), the scaffold plate was set in a conventional
centrifugal machine placed in an incubator. The experimental
results show that the tendency of cells to align the longitudinal
direction of the striped pattern is strengthened by the hysteresisof the exposure to the parallel tangential force field. The
tendency is reset after the rounded shape at the division of cell.

[10] H. Hino, S. Hashimoto and F. Sato, “Effect of Micro
Ridges on Orientation of Cultured Cell”, Journal of
Systemics Cybernetics and Informatics, Vol. 12, No. 3,
2014, pp. 47–53.
[11] Y. Endo, S. Hashimoto and T. Tamura, “Alignment andDeformation of MC3T3-E1 Cultured on Micro Striped
Pattern after Stimulation of Tangential Force Field”, Proc.
23rd World Multi-Conference on Systemics Cybernetics
and Informatics, Vol. 2, 2019, pp. 76–81.
[12] T. Tamura, S. Hashimoto, H. Hino, T. Ito and Y. Endo,“Tracings of Orientation of Cell on Scaffold with Micro
Striped Pattern after Stimulation of Vertical Excess Gravity”,
Proc. 22nd World Multi-Conference on Systemics
Cybernetics and Informatics, Vol. 2, 2018, pp. 31–36.
[13] T. Tamura, H. Hino, S. Hashimoto, H. Sugimoto and Y.
Takahashi, “Cell Behavior After Stimulation of Excess
Gravity”, Proc. 21st World Multi-Conference on
Systemics Cybernetics and Informatics, Vol. 2, 2017, pp.
263–268.
[14] J. Eckert, J.J.W.A. van Loon, L. M. Eng and T. Schmidt,“Hypergravity Affects Cell Traction Forces of Fibroblasts”,
bioRxiv, 2020, https://doi.org/10.1101/2020.03.30.015958[15] P. Sawadkar, D. Player, L. Bozec and V. Mudera, “TheMechanobiology of Tendon Fibroblasts under Static andUniaxial Cyclic Load in a 3D Tissue Engineered Model
Mimicking Native Extracellular Matrix”, Journal of Tissue
Engineering and Regenerative Medicine, Vol. 14, No. 1,
2019, https://doi.org/10.1002/term.2975[16] T. Tominari, R. Ichimaru, K. Taniguchi, A. Yumoto, M.
Shirakawa, C. Matsumoto, K. Watanabe, M. Hirata, Y. Itoh,D. Shiba, C. Miyaura and M. Inada, “Hypergravity andMicrogravity Exhibited Reversal Effects on the Bone andMuscle Mass in Mice”, Scientific Reports, Vol. 9, No.
6614, 2019.
[17] K.W. Aguilar‐Agon, A.J. Capel, N.R.W. Martin, D.J. Playerand M.P. Lewis, “Mechanical Loading Stimulates
Hypertrophy in Tissue‐Engineered Skeletal Muscle:
Molecular and Phenotypic Responses”, Journal of Cellular
Physiology, Vol. 234, No. 12, 2019, pp. 23547–23558.
[18] D. Frank, A. Cser, B. Kolarovszki, N. Farkas, A. Miseta
and T. Nagy, “Mechanical Stress Alters Protein O-GlcNAc
in Human Periodontal Ligament Cells”, Journal of Cellular
and Molecular Medicine, Vol. 23, No. 9, 2019, DOI:
10.1111/jcmm.14509[19] H. Kanzaki, M. Chiba, Y. Shimizu and H. Mitani,“Periodontal Ligament Cells under Mechanical Stress Induce
Osteoclastogenesis by Receptor Activator of Nuclear Factor
κB Ligand Up-Regulation via Prostaglandin E2 Synthesis”,
Journal of Bone and Mineral Research, Vol. 17, No. 2,
2009, https://doi.org/10.1359/jbmr.2002.17.2.210[20] S.H. Chang, D. Mori, H. Kobayashi, Y. Mori, H. Nakamoto,
K. Okada, Y. Taniguchi, S. Sugita, F. Yano, U. Chung, J.
Kim-Kaneyama, M. Yanagita, A. Economides, E. Canalis, D.
Chen, S. Tanaka and T. Saito, “Excessive Mechanical
Loading
Promotes
Osteoarthritis
throughtheGremlin-1–NF-κB Pathway”, Nature Communications, Vol.
10, No. 1442, 2019, pp. 1–13.

ACKNOWLEDGMENT
The authors thank to Mr. Takuya Tamura for his assistance ofthe experiment.

REFERENCES[1] H. Hino, S. Hashimoto, Y. Shinozaki, H. Sugimoto and Y.
Takahashi, “Effect of Flow on Cultured Cell at Micro-patternof Ridge Lines”, Journal of Systemics Cybernetics andInformatics, Vol. 15, No. 5, 2017, pp. 1–7.
[2] S. Hashimoto, Y. Takahashi, K. Kakishima and Y. Takiguchi,“Slit between Micro Machined Plates for Observation of
Passing Cell: Deformation and Velocity”, Proc. 22nd World
Multi-Conference on Systemics Cybernetics andInformatics, Vol. 2, 2018, pp. 102–107.
[3] A. Mizoi, Y. Takahashi, H. Hino, S. Hashimoto and T.
Yasuda, “Deformation of Cell Passing through Micro Slit”,
Proc. 19th World Multi-Conference on Systemics
Cybernetics and Informatics, Vol. 2, 2015, pp. 270–275.
[4] H. Sugimoto, H. Hino, Y. Takahashi and S. Hashimoto,“Effect of Surface Morphology of Scaffold with Lines ofMicro Ridges on Deformation of Cells”, Proc. 20th World
Multi-Conference on Systemics Cybernetics andInformatics, Vol. 2, 2016, pp. 135–140.
[5] H. Hino, S. Hashimoto, S. Nishino, H. Nakajima, Y.
Takahashi and H. Sugimoto, “Behavior of Cell on Vibrating
Micro Ridges”, Journal of Systemics Cybernetics andInformatics, Vol. 13, No. 3, 2015, pp. 9–16.
[6] S. Hashimoto, H. Hino, T. Yamamoto, T. Tamura and K. Abe,
“Design of Surface Electrode for Measurement of Electric
Impedance of Arrangement of Cells Oriented on Micro
Striped Pattern”, Proc. 22nd World Multi-Conference on
Systemics Cybernetics and Informatics, Vol. 2, 2018, pp.
114–119.
[7] H. Hino, H. Nakajima, S. Hashimoto, N. Wakuri, Y.
Takahashi and T. Yasuda, “Effect of Electric Stimulation on
Differentiation and Hypertrophy of Fat Precursor Cells”,
Proc. 19th World Multi-Conference on Systemics
Cybernetics and Informatics, Vol. 2, 2015, pp. 252–257.
[8] Y. Takahashi, S. Hashimoto, H. Hino, A. Mizoi and N.
Noguchi, “Micro Groove for Trapping of Flowing Cell”,
Journal of Systemics Cybernetics and Informatics, Vol.
13, No. 3, 2015, pp. 1–8.
[9] T. Tamura, H. Hino and S. Hashimoto, “Behavior of Cell
Cultured on Micro Striped Pattern after Stimulation of
Excess Gravity”, Proc. 22nd World Multi-Conference on
Systemics Cybernetics and Informatics, Vol. 2, 2018, pp.
25–30.

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