Movement of Myoblast Flowing Through Electric Field Perpendicular to Flow Channel

2021 ◽  
Author(s):  
Shigehiro Hashimoto ◽  
Kiyoshi Yoshinaka
Keyword(s):  
Electric Field ◽  
Flow Direction ◽  
Reference Electrode ◽  
Major Axis ◽  
Surface Electrodes ◽  
Myoblast Cell Line ◽  
Main Flow Direction ◽  
Flat Edge ◽  
Myoblast Cell ◽  
Mouse Myoblast

Abstract The sorting technology with little invasion to cells would be applied to regenerative medicine and diagnosis. In this study, dielectrophoresis is focused on. The dielectrophoretic effect on the flowing myoblasts was maximized by adjusting several parameters: the shape of the electrodes, the amplitude and frequency of the alternating current. The suspension of C2C12 (mouse myoblast cell line) was injected into the channel, and the movement of each flowing cell was analyzed at the microscopic movie image. A pair of titanium-coated (200 nm thick) asymmetric surface electrodes (a triangular electrode with a tip angle of 0.35 rad and a rectangular reference electrode with a flat edge) was manufactured by photolithography technique. With the alternating square cyclic wave at the frequency of 3 MHz and the amplitude of current of ± 7.5 mA, 70 μm movement along the electric field (perpendicular to the main flow direction) of the cell was obtained. The movement along the electric field is governed by several parameters of the cell: the diameter, the deformation ratio, and the direction of the major axis. The method can be applied to cell sorting.

scholarly journals How Does a Cell Change Flow Direction Due to a Micro Groove?

2021 ◽  
Vol 19 (8) ◽  
pp. 164-181
Author(s):  
Shigehiro Hashimoto ◽  
Taketo Matsumoto ◽  
Shogo Uehara
Keyword(s):  
Flow Direction ◽  
Longitudinal Axis ◽  
Main Flow ◽  
Myoblast Cell Line ◽  
A Cell ◽  
Micro Flow ◽  
Main Flow Direction ◽  
Myoblast Cell ◽  
Mouse Myoblast

The change in direction of a cell flowing over an oblique micro groove has been analyzed in vitro. The micro flow-channel (0.05 mm height x 1 mm width x 25 mm length) with oblique micro grooves (4.5 μm depth) was manufactured on a polydimethylsiloxane (PDMS) disk by the micromachining technique. The angle between the main flow direction and the longitudinal axis of the groove is 45 degrees. The effect of variation of the groove width (0.03 mm, 0.04 mm, and 0.05 mm) was studied. Myoblasts (C2C12: mouse myoblast cell line) were used in the test. The main flow velocity (0.02 mm/s < vx < 0.23 mm/s) of the medium was controlled by the pressure difference between the inlet and the outlet. The shape of each flowing cell was tracked on a movie recorded by the camera attached to the eyepiece of the microscope. The experimental results show that the change of the direction of each cell by each groove depends on the shape of the cell, which depends on both the shape of the cell and the width of the groove.

Oblique Micro Grooves on Bottom Wall of Flow Channel to Sort Cells

2020 ◽  
Author(s):  
Shigehiro Hashimoto
Keyword(s):  
Flow Direction ◽  
Longitudinal Axis ◽  
Flow Channel ◽  
Bottom Surface ◽  
Rectangular Shape ◽  
Myoblast Cell Line ◽  
Micro Flow ◽  
Myoblast Cell ◽  
Mouse Myoblast

Abstract The movement of a flowing cell near the oblique micro groove on the bottom surface in the micro flow channel has been measured to sort biological cells in vitro. The micro groove of the rectangular shape (4.5 μm depth, and 0.2 mm length) was fabricated on the polydimethylsiloxane (PDMS) disk by the photolithography technique. The angle between the flow direction and the longitudinal axis of the groove is 45 degree. Variation has been made on the width (0.03 mm &lt; w &lt; 0.05 mm) of the groove. A rectangular flow channel (0.05 mm height × 1 mm width × 25 mm length) has been constructed between two transparent PDMS disks. C2C12 (mouse myoblast cell line) was used in the test. A flow velocity (0.1 mm/s &lt; vx &lt; 2.4 mm/s) of the suspension of cells was controlled by the pressure difference between the inlet and the outlet. The shifted distance of each cell along the oblique groove depends on the diameter of the cell. The malnourished cell with the different density can be distinguished by the shifted distance according to the velocity of the cell.

Movement of Cell Flowing Over Oblique Micro Grooves in Flow Channel

2021 ◽  
Author(s):  
Shigehiro Hashimoto ◽  
Hiroki Yonezawa
Keyword(s):  
Flow Direction ◽  
Longitudinal Axis ◽  
Flow Channel ◽  
Main Flow ◽  
Flow Test ◽  
Micro Flow ◽  
Main Flow Direction ◽  
Mouse Myoblast ◽  
Made In

Abstract A micro flow-channel with bottom-microgrooves has been manufactured by photolithography technique for cell sorting. The movement of each cell passing over microgrooves has been analyzed in relation to cell deformation and alignment in vitro. The flow path (height 0.05 mm × width 1 mm × length 25 mm) between the two transparent PDMS disks has rectangular microgrooves (4.5 μm deep, 0.2 mm long) on the bottom. Variations are made in groove widths (0.03 mm, 0.04 mm, and 0.05 mm). The angle between the flow direction and the longitudinal axis of the groove is 45 degrees. Myoblasts (C2C12: mouse myoblast line) were used in the flow test. The main flow velocity of the medium (0.02 mm/s &lt; vx &lt; 0.23 mm/s) was controlled by the pressure difference between the inlet and the outlet. The shape of each flowing cell was tracked in a movie recorded by a camera attached to the eyepiece of the microscope. Experimental results show that the movement perpendicular to the main flow direction in the micro-groove can distinguish cells in relation to smaller deformations and larger alignment changes.

scholarly journals PFN2a Suppresses C2C12 Myogenic Development by Inhibiting Proliferation and Promoting Apoptosis via the p53 Pathway

Cells ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 959 ◽  
Author(s):  
Huaqin Li ◽  
Lianjie Hou ◽  
Yu Zhang ◽  
Fangyi Jiang ◽  
Yifan Zhu ◽  
...  
Keyword(s):  
Myogenic Differentiation ◽  
Inhibitory Effect ◽  
Cancer Growth ◽  
Actin Monomer ◽  
P53 Pathway ◽  
Histone Deacetylase 1 ◽  
Myoblast Cell Line ◽  
Myoblast Cell ◽  
Body Energy ◽  
Mouse Myoblast

Skeletal muscle plays a crucial role in physical activity and in regulating body energy and protein balance. Myoblast proliferation, differentiation, and apoptosis are indispensable processes for myoblast myogenesis. Profilin 2a (PFN2a) is a ubiquitous actin monomer-binding protein and promotes lung cancer growth and metastasis through suppressing the nuclear localization of histone deacetylase 1 (HDAC1). However, how PFN2a regulates myoblast myogenic development is still not clear. We constructed a C2C12 mouse myoblast cell line overexpressing PFN2a. The CRISPR/Cas9 system was used to study the function of PFN2a in C2C12 myogenic development. We find that PFN2a suppresses proliferation and promotes apoptosis and consequentially downregulates C2C12 myogenic development. The suppression of PFN2a also decreases the amount of HDAC1 in the nucleus and increases the protein level of p53 during C2C12 myogenic development. Therefore, we propose that PFN2a suppresses C2C12 myogenic development via the p53 pathway. Si-p53 (siRNA-p53) reverses the PFN2a inhibitory effect on C2C12 proliferation and the PFN2a promotion effect on C2C12 apoptosis, and then attenuates the suppression of PFN2a on myogenic differentiation. Our results expand understanding of PFN2a regulatory mechanisms in myogenic development and suggest potential therapeutic targets for muscle atrophy-related diseases.

scholarly journals Differential localization of the mRNA of the M and B isoforms of creatine kinase in myoblasts

1995 ◽  
Vol 308 (2) ◽  
pp. 599-605 ◽  
Author(s):  
I A Wilson ◽  
K M Brindle ◽  
A M Fulton
Keyword(s):  
Creatine Kinase ◽  
Cell Line ◽  
Cell Periphery ◽  
Energy Demands ◽  
Enzyme Isoforms ◽  
Myoblast Cell Line ◽  
Temporal Aspect ◽  
Myoblast Cell ◽  
Mouse Myoblast

Creatine kinase (CK) plays an important role in buffering ATP and ADP levels in tissues which have intermittently high and fluctuating energy demands, such as skeletal muscle. This buffering function has a spatial, as well as a temporal aspect, which is dependent on the localization of different enzyme isoforms within the cell. We show here, by in situ hybridization, that the mRNAs for the cytoplasmic isoforms of CK are differentially localized in a mouse myoblast cell line (C2C12). The mRNA for the M form is localized at the cell periphery, while that for the B form is localized in the perinuclear region. Deletion of segments of the 3′ untranslated regions of these mRNAs or swapping of these segments between the mRNAs for the two isoforms demonstrated that localization signals lie within these regions. Localization appears to be tissue-specific, since both the M and B mRNAs were distributed uniformly over the cytoplasm in a non-muscle cell line. These results, in conjunction with other studies which have shown that mRNA localization can lead to co-localization of the encoded protein, suggest that the localization of the mRNAs for the cytoplasmic isoforms of CK may be involved in the localization of the enzymes themselves.

Control of intracellular creatine concentration in a mouse myoblast cell line

1993 ◽  
Vol 21 (4) ◽  
pp. 441S-441S ◽  
Author(s):  
J. E. ODOOM ◽  
G. J. KEMP ◽  
G. K. RADDA
Keyword(s):  
Cell Line ◽  
Myoblast Cell Line ◽  
Myoblast Cell ◽  
Mouse Myoblast

scholarly journals Zebrafish Melanophores Suggest Novel Functions of Cell Chirality in Tissue Formation

Symmetry ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 130
Author(s):  
Hiroaki Yamanaka ◽  
Shigeru Kondo
Keyword(s):  
Cell Line ◽  
C2c12 Cells ◽  
Tissue Formation ◽  
Spiral Pattern ◽  
Leftward Bias ◽  
Asymmetric Behavior ◽  
Myoblast Cell Line ◽  
Culture Environment ◽  
Myoblast Cell ◽  
Mouse Myoblast

Several types of cells show left–right asymmetric behavior, unidirectional rotation, or spiral movements. For example, neutrophil-like differentiated HL60 (dHL60) cells show leftward bias in response to chemoattractant. Neurons extend neurites, creating a clockwise spiral. Platelet cells shows unidirectional spiral arrangements of actin fibers. In the microfabricated culture environment, groups of C2C12 cells (mouse myoblast cell line) were autonomously aligned in a counter-clockwise spiral pattern, and isolated C2C12 cells showed unidirectional spiral pattern of the actin skeleton. This biased directionality suggested that these cells have inherent cell chirality. In addition to these cells, we recently found that melanophores of zebrafish also have an intrinsic cellular chirality that was shown by their counter-clockwise self-rotation. Although this cell chirality is obvious, the function of the cell chirality is still unclear. In this review, we compare the cell chirality of melanophores of zebrafish with other cell chirality and consider the function of cell chirality in morphogenesis.

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