Maintenance of contractile function of isolated airway smooth muscle after cryopreservation.
Abstract: Isolated human airway smooth muscle (ASM) tissue contractility studies are essential for understanding the role of ASM in respiratory disease, but limited availability and cost render storage options necessary for optimal use. However, to our knowledge, no comprehensive study of cryopreservation protocols for isolated ASM has been performed to date. We tested several cryostorage protocols on equine trachealis ASM using different cryostorage media [1.8 M dimethyl sulfoxide and fetal bovine serum (FBS) or Krebs-Henseleit (KH)] and different degrees of dissection (with or without epithelium and connective tissues attached) before storage. We measured methacholine (MCh), histamine, and isoproterenol (Iso) dose-responses and electrical field stimulation (EFS) and MCh force-velocity curves. We confirmed our findings in human trachealis ASM stored undissected in FBS. Maximal stress response to MCh was decreased more in dissected than undissected equine tissues. EFS force was decreased in all equine but not in human cryostored tissues. Furthermore, in human cryostored tissues, EFS maximal shortening velocity was decreased, and Iso response was potentiated after cryostorage. Overnight incubation with 0.5 or 10% FBS did not recover contractility in the equine tissues but potentiated Iso response. Overnight incubation with 10% FBS in human tissues showed maximal stress recovery and maintenance of other contractile parameters. ASM tissues can be cryostored while maintaining most contractile function. We propose an optimal protocol for cryostorage of ASM as undissected tissues in FBS or KH solution followed by dissection of the ASM bundles and a 24-h incubation with 10% FBS before mechanics measurements.
Publication Date: 2018-08-09 PubMed ID: 30091377DOI: 10.1152/ajplung.00064.2018Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research focuses on perfecting methods to cryostore isolated human airway smooth muscle tissues, important for understanding respiratory diseases. Researchers tested multiple cryostorage protocols and present one that allows the tissues to maintain most of their contractile function.
Overview of Research
The study conducted a comprehensive exploration of several protocols for freezing and storing isolated airway smooth muscle (ASM), focusing on human and equine trachealis ASM. These muscles are integral to understanding various respiratory diseases, hence the importance of preservation for study. Lack of availability and high cost called for robust storage options.
Cryostorage Protocols
- The research team tested different cryostorage media: 1.8 M dimethyl sulfoxide with fetal bovine serum (FBS) or Krebs-Henseleit solution (KH).
- The tissues were also tested at differing levels of dissection, either with epithelium and connective tissues attached or without.
- Contractility responses to drugs (methacholine, histamine, and isoproterenol) and electrical field stimulation were measured before and after cryostorage.
Results of the Cryostorage
- Evidence showed that maximal stress response to methacholine decreased more in dissected equine tissues than in intact ones.
- EFS force decreased in all equine cryostored tissues, whereas the human tissues showed no similar decline.
- Maximal shortening velocity due to EFS was reduced in human cryostored tissues, and isoproterenol response was potentiated after cryostorage.
- Overnight incubation with 0.5 or 10% FBS did not recover contractility in equine tissues, but it did potentiate the Iso response.
- In human tissues, overnight incubation with 10% FBS led to maximal stress recovery and maintenance of other contractile parameters.
Recommended Protocol
Based on their findings, the researchers proposed an optimal protocol for cryostorage. This involves storing ASM as undissected tissues in FBS or KH solution, followed by dissection of the ASM bundles and a 24-hour incubation with 10% FBS before conducting measurements. This protocol enables the isolated ASM tissues to retain most of their contractile function even after cryostorage. The results of this research can be beneficial in numerous future studies involving the use of human airway smooth muscle tissues.
Cite This Article
APA
Ijpma G, Liang CQ, Kachmar L, Panariti A, Benedetti A, Lavoie JP, Lauzon AM.
(2018).
Maintenance of contractile function of isolated airway smooth muscle after cryopreservation.
Am J Physiol Lung Cell Mol Physiol, 315(5), L724-L733.
https://doi.org/10.1152/ajplung.00064.2018 Publication
Researcher Affiliations
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Meakins-Christie Laboratories, Research Institute of the McGill University Health Centre , Montreal, Q , Canada.
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Meakins-Christie Laboratories, Research Institute of the McGill University Health Centre , Montreal, Q , Canada.
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Meakins-Christie Laboratories, Research Institute of the McGill University Health Centre , Montreal, Q , Canada.
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Department of Epidemiology, Biostatistics, and Occupational Health, McGill University , Montreal, Q , Canada.
- Respiratory Epidemiology and Clinical Research Unit, McGill University Health Centre , Montreal, Q , Canada.
- Faculty of Veterinary Medicine, Department of Clinical Sciences, University of Montreal , Saint-Hyacinthe, Q , Canada.
- Department of Medicine, McGill University , Montreal, Q , Canada.
- Meakins-Christie Laboratories, Research Institute of the McGill University Health Centre , Montreal, Q , Canada.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryoprotective Agents / chemistry
- Dimethyl Sulfoxide / chemistry
- Histamine / chemistry
- Horses
- Methacholine Chloride / chemistry
- Muscle Contraction / physiology
- Muscle, Smooth / cytology
- Muscle, Smooth / physiology
- Trachea / cytology
- Trachea / physiology
Grant Funding
- Canadian Institutes of Health Research
Citations
This article has been cited 1 times.- Ram-Mohan S, Bai Y, Schaible N, Ehrlicher AJ, Cook DP, Suki B, Stoltz DA, Solway J, Ai X, Krishnan R. Tissue traction microscopy to quantify muscle contraction within precision-cut lung slices.. Am J Physiol Lung Cell Mol Physiol 2020 Feb 1;318(2):L323-L330.
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