التفاصيل البيبلوغرافية
| العنوان: |
A method for isolating contractile smooth muscle cells from cryopreserved tissue. |
| المؤلفون: |
Schultz MLC; Department of Biomedical Engineering, McGill University, Montreal, Quebec, Canada.; Research Institute of the McGill University Health Centre, Meakins-Christie Laboratories, Montreal, Quebec, Canada., Seth P; Department of Biomedical Engineering, McGill University, Montreal, Quebec, Canada.; Research Institute of the McGill University Health Centre, Meakins-Christie Laboratories, Montreal, Quebec, Canada., Kachmar L; Research Institute of the McGill University Health Centre, Meakins-Christie Laboratories, Montreal, Quebec, Canada., Ijpma G; Research Institute of the McGill University Health Centre, Meakins-Christie Laboratories, Montreal, Quebec, Canada., Lauzon AM; Department of Biomedical Engineering, McGill University, Montreal, Quebec, Canada.; Research Institute of the McGill University Health Centre, Meakins-Christie Laboratories, Montreal, Quebec, Canada.; Department of Medicine, McGill University, Montreal, Quebec, Canada. |
| المصدر: |
American journal of physiology. Cell physiology [Am J Physiol Cell Physiol] 2024 Mar 01; Vol. 326 (3), pp. C990-C998. Date of Electronic Publication: 2024 Feb 05. |
| نوع المنشور: |
Journal Article |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: American Physiological Society Country of Publication: United States NLM ID: 100901225 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1522-1563 (Electronic) Linking ISSN: 03636143 NLM ISO Abbreviation: Am J Physiol Cell Physiol Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: Bethesda, Md. : American Physiological Society, |
| مواضيع طبية MeSH: |
Muscle Contraction* , Myocytes, Smooth Muscle*/metabolism, Muscle, Smooth/metabolism ; Phenotype ; Contractile Proteins/genetics ; Contractile Proteins/metabolism ; Cells, Cultured ; Cell Differentiation/genetics |
| مستخلص: |
Multiple techniques have been developed to isolate contractile smooth muscle cells (SMCs) from tissues with varying degrees of success. However, most of these approaches rely on obtaining fresh tissue, which poses logistical challenges. In the present study, we introduce a novel protocol for isolating contractile SMCs from cryopreserved smooth muscle (SM) tissue, thereby enhancing experimental efficiency. This protocol yields abundant viable, spindle-shaped, contractile SMCs that closely resemble those obtained from fresh samples. By analyzing the expression of contractile proteins, we demonstrate that both the isolated SMCs from cryopreserved tissue represent more accurately fresh SM tissue compared with cultured SMCs. Moreover, we demonstrate the importance of a brief incubation step of the tissue in culture medium before cell dissociation to achieve contractile SMCs. Finally, we provide a concise overview of our protocol optimization efforts, along with a summary of previously published methods, which could be valuable for the development of similar protocols for other species. NEW & NOTEWORTHY We report a successful protocol development for isolating contractile smooth muscle cells (SMCs) from cryopreserved tissue reducing the reliance on fresh tissues and providing a readily available source of contractile SMCs. Our findings suggest that SMCs isolated using our protocol maintain their phenotype better compared with cultured SMCs. This preservation of the cellular characteristics, including the expression of key contractile proteins, makes these cells more representative of fresh SM tissue. |
| معلومات مُعتمدة: |
RGPIN#05054-2015 Canadian Government | Natural Sciences and Engineering Research Council of Canada (NSERC); PJT156369 Canadian Government | Canadian Institutes of Health Research (IRSC) |
| فهرسة مساهمة: |
Keywords: cell contractility; cell shortening; single-cell studies; smooth muscle cells |
| المشرفين على المادة: |
0 (Contractile Proteins) |
| تواريخ الأحداث: |
Date Created: 20240205 Date Completed: 20240306 Latest Revision: 20240306 |
| رمز التحديث: |
20240306 |
| DOI: |
10.1152/ajpcell.00442.2023 |
| PMID: |
38314725 |
| قاعدة البيانات: |
MEDLINE |
