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Overview
This video protocol demonstrates the dissociation of adult mouse lung tissue using the gentleMACS dissociator. The procedure is designed to produce a high volume of viable, culturable cells for further applications.
Key Study Components
Area of Science
- Cell dissociation techniques
- Single cell suspension preparation
- Mouse lung tissue analysis
Background
- Dissociating cells from tissues requires specific parameters.
- The gentleMACS dissociator provides an optimized protocol.
- Collagenase D and DNase are used in the dissociation process.
- Filtered cell suspensions are prepared for further applications.
Purpose of Study
- To demonstrate a reproducible method for preparing single cell suspensions.
- To facilitate the study of leukocytes and endothelial cells.
- To provide a practical protocol for researchers in the field.
Methods Used
- Use of gentleMACS dissociator for tissue agitation.
- Application of collagenase D and DNase during dissociation.
- Filtration of cell suspensions through a 70 micron cell strainer.
- Centrifugation and suspension of cells for further applications.
Main Results
- Successful dissociation of mouse lung tissue into single cell suspensions.
- High volume of viable, culturable cells obtained.
- Protocol demonstrated reproducibility and efficiency.
- Preparation suitable for downstream applications.
Conclusions
- The gentleMACS dissociator is effective for lung tissue dissociation.
- This method can be applied to various tissue types.
- Provides a valuable tool for researchers studying lung tissue.
What is the gentleMACS dissociator?
It is a device used to dissociate tissues into single cell suspensions efficiently.
What type of tissue is demonstrated in this protocol?
The protocol demonstrates the dissociation of adult mouse lung tissue.
What enzymes are used in the dissociation process?
Collagenase D and DNase are used to facilitate the dissociation.
How are the cell suspensions filtered?
Cell suspensions are filtered using a 70 micron cell strainer.
What applications can the prepared cell suspensions be used for?
They can be used for studying leukocytes and endothelial cells.
Is the protocol reproducible?
Yes, the protocol is designed to be reproducible and efficient.
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