ISOLATION AND OPTIMIZATION OF MICE SKELETAL MUSCLE SATELLITE CELLS USING PREPLATING METHOD AND CULTURE MEDIA SUBSTITUTION

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BAKHTIARI NUREDIN

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Journal Paper

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Journal: Physiology and Pharmacology Year:2016 | Volume:20 | Issue:1 Page(s): 63-73

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Information Journal Paper

Title

ISOLATION AND OPTIMIZATION OF MICE SKELETAL MUSCLE SATELLITE CELLS USING PREPLATING METHOD AND CULTURE MEDIA SUBSTITUTION

Author(s)

BAKHTIARI NUREDIN | Issue Writer Certificate

Keywords

SKELETAL MUSCLE

SATELLITE CELLS

PREPLATE TECHNIQUE

PAX7

Abstract

Introduction: SATELLITE CELLS are known as the main regenerative cell type in SKELETAL MUSCLEs. Our study established a modified digestion and preplating method for the isolation of slow or weak adherent cells for the enrichment of SATELLITE CELLS. Low-survival rate of these primary stem cells prompted us to address whether cell culture medium substitution might change cell viability status.Methods: SKELETAL MUSCLE from 10-day-NMRI mice was gently isolated, dissected and digested by collagenase type I, IV and dispases. The isolated cells were verified by cellular (immunocytochemistry and flow-cytometry) and molecular (real-time PCR) techniques and the results were compared with sub-cultured cells (non-preplated cells) to determine the efficiency of preplating technique as a common isolating procedure of SATELLITE CELLS. All data were analyzed using SPSS 16 and One Away ANOVA test.Results: The isolated cells exhibited a close gene expression pattern with SATELLITE CELLS for self-renewal and fusion phases. The findings revealed that PAX7 as a self-renewal marker was expressed ~ 201.4 times higher than sub-cultured-group. Moreover, the findings obviously indicated that substitution of a-MEM to DMEM cell culture medium improves the survival rates of the cells.Conclusion: Our results recommend that preplating technique is a useful procedure for the isolation of SATELLITE CELLS. In addition, it seems that substitution of culture medium paves the way for investigators to seek various therapeutic methods for SKELETAL MUSCLE-related disorders such as SKELETAL MUSCLE atrophy (SMA), amyotrophic lateral sclerosis (ALS), sarcopenia, diabetes and aging.

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APA: Copy

BAKHTIARI, NUREDIN. (2016). ISOLATION AND OPTIMIZATION OF MICE SKELETAL MUSCLE SATELLITE CELLS USING PREPLATING METHOD AND CULTURE MEDIA SUBSTITUTION. PHYSIOLOGY AND PHARMACOLOGY, 20(1), 63-73. SID. https://sid.ir/paper/696103/en

Vancouver: Copy

BAKHTIARI NUREDIN. ISOLATION AND OPTIMIZATION OF MICE SKELETAL MUSCLE SATELLITE CELLS USING PREPLATING METHOD AND CULTURE MEDIA SUBSTITUTION. PHYSIOLOGY AND PHARMACOLOGY[Internet]. 2016;20(1):63-73. Available from: https://sid.ir/paper/696103/en

IEEE: Copy

NUREDIN BAKHTIARI, “ISOLATION AND OPTIMIZATION OF MICE SKELETAL MUSCLE SATELLITE CELLS USING PREPLATING METHOD AND CULTURE MEDIA SUBSTITUTION,” PHYSIOLOGY AND PHARMACOLOGY, vol. 20, no. 1, pp. 63–73, 2016, [Online]. Available: https://sid.ir/paper/696103/en

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