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311155

Optimization of Single Myofiber Isolation Protocol from the Flexor Digitorum Brevis Muscle to Study Satellite Cells

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Last updated: 04 Jan 2025

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Basic Research

Abstract

Background: Satellite cells, which are located between the basal lamina and sarcolemma of the myofiber, have a tremendous ability to repair damaged skeletal muscle. Although they remain quiescent in their niche, they activate rapidly following injury; understanding the mechanisms underlying this activation process has the potential to open new avenues for muscle regenerative therapies. The flexor digitorum brevis muscle “FDB" is one of the best candidates to study satellite cell physiology in the unperturbed myofiber, as it is composed of relatively short myofibers attached to the muscle tendon in an oblique manner, enabling a high rate of intact fiber isolation compared to muscles with longer fibers. Aim: To optimize the preparation process to avoid maldigestion that renders broken and contracted myofibers not suitable for studying quiescent satellite cells and the subsequent activation processes. Material and Methods: FDB muscles from both sides from C57BL/6 male mice were used. The muscles were digested with collagenase digestion solution then cultured in myofiber culture medium in low CO2 incubators at 370C. Results: This study revealed that a short incubation period in the digestion solution of two hours is optimum for harvesting the largest number of viable myofiber. Dishes coated with laminin were used to allow the attachment of the single myofibers and further facilitate the immunofluorescence staining process. Conclusion: The used method provides a better tool to isolate the single myofibers from the FDB muscle and hence better analysis of satellite cell activation.
 

DOI

10.21608/scumj.2023.311155

Keywords

satellite cells, FDB, single myofiber, activation

Authors

First Name

Ahmed

Last Name

Ahmed

MiddleName

-

Affiliation

Department of Anatomy, Faculty of Medicine, Suez Canal University, Ismailia, Egypt, \University of Minnesota Twin Cities, Minneapolis, Minnesota, USA

Email

ahmedsabershams@gmail.com

City

-

Orcid

-

First Name

Hoda H.

Last Name

Hussein

MiddleName

-

Affiliation

Department of Anatomy, Faculty of Medicine, Suez Canal University, Ismailia, Egypt

Email

-

City

-

Orcid

-

First Name

Omayma M.

Last Name

Mahmoud

MiddleName

-

Affiliation

Department of Anatomy, Faculty of Medicine, Suez Canal University, Ismailia, Egypt

Email

-

City

-

Orcid

-

First Name

Gamal

Last Name

Abdel-Alrahman

MiddleName

-

Affiliation

Department of Anatomy, Faculty of Medicine, Suez Canal University, Ismailia, Egypt

Email

-

City

-

Orcid

-

First Name

Michael

Last Name

Kyba

MiddleName

-

Affiliation

University of Minnesota Twin Cities, Minneapolis, Minnesota, USA

Email

mkx012@hotmail.com

City

-

Orcid

-

First Name

Shimaa A.

Last Name

Fareed

MiddleName

-

Affiliation

Department of Anatomy, Faculty of Medicine, Suez Canal University, Ismailia, Egypt

Email

-

City

-

Orcid

-

Volume

26

Article Issue

7

Related Issue

42539

Issue Date

2023-07-01

Receive Date

2023-08-04

Publish Date

2023-07-01

Page Start

1

Page End

9

Print ISSN

1110-6999

Online ISSN

2090-2581

Article File

SCUMJ_Volume 26_Issue 7_Pages 1-9.pdf

PDF . 475.3kB

Link

https://scumj.journals.ekb.eg/article_311155.html

Detail API

https://scumj.journals.ekb.eg/service?article_code=311155

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Type

Original Article

Type Code

938

Publication Type

Journal

Publication Title

Suez Canal University Medical Journal

Publication Link

https://scumj.journals.ekb.eg/

MainTitle

Optimization of Single Myofiber Isolation Protocol from the Flexor Digitorum Brevis Muscle to Study Satellite Cells

Details

Type

Article

Created At

24 Dec 2024