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Pepsin can be used to subculture viable mammary epithelial cells
In Vitro
|September 1, 1983
Abstract:
Normal mouse mammary epithelial cells in primary culture can be passaged as viable single cells using 0.5 to 1.0 mg/ml pepsin in Hanks' salt solution. After 5 min the pepsin treatment preferentially removes fibroblasts, leaving a monolayer of purified epithelial cells that can be removed by pipetting and transferred to new culture vessels or injected into animals.
Insights
Researchers developed a method to isolate pure mouse mammary epithelial cells. Pepsin treatment effectively removes fibroblasts, enabling cell transfer for further research or animal studies.
Area of Science:
- Cell biology
- Biotechnology
- Mammalian cell culture
Background:
- Primary cell cultures are essential for biological research.
- Mammary epithelial cells are crucial for studying mammary gland development and diseases.
- Current methods for isolating pure epithelial cells can be challenging and time-consuming.
Purpose of the Study:
- To develop an efficient method for isolating pure mouse mammary epithelial cells from primary cultures.
- To assess the efficacy of pepsin treatment in removing contaminating fibroblasts.
Main Methods:
- Primary mouse mammary epithelial cells were cultured.
- Cells were treated with pepsin (0.5-1.0 mg/ml) in Hanks' salt solution for 5 minutes.
- Epithelial cell monolayers were purified and collected by pipetting.
Main Results:
- Pepsin treatment effectively removed fibroblasts, a common contaminant in primary cell cultures.
- A monolayer of purified epithelial cells was obtained.
- The isolated epithelial cells remained viable for subsequent transfer or injection.
Conclusions:
- Pepsin treatment offers a rapid and effective method for purifying mouse mammary epithelial cells.
- This technique facilitates the isolation of viable epithelial cells for downstream applications in research and animal models.