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Facilitation of mouse skin-derived precursor growth and yield by optimizing plating density
Yiming Li1,2, Lidan Xiong1, Jie Tang1
1Department of Dermatology and Venerology, West China Hospital, Sichuan University, 37 Guoxue Alley, Chengdu, Sichuan Province 610041, China.
Open Life Sciences
|December 30, 2021
Summary
Determining optimal plating density for mouse skin-derived precursors (mSKPs) is crucial. The ideal density (2.5-5 × 105 cells/mL) enhances mSKP sphere growth and yield from adherent cell colonies.
Area of Science:
- Stem Cell Biology
- Dermatology
- Tissue Engineering
Background:
- Skin-derived precursors (SKPs) are multipotent stem cells with therapeutic potential.
- Existing SKP culture methods lack optimization regarding plating density.
- Understanding SKP proliferation and differentiation is key for regenerative medicine.
Purpose of the Study:
- To identify the optimal plating density for mouse SKP (mSKP) culture.
- To investigate the relationship between adherent cell colonies and mSKP sphere formation.
- To characterize the differentiation potential of adherent cell colonies.
Main Methods:
- Culturing mouse SKPs at six different plating densities in two flask types.
- Assessing mSKP sphere number, diameter, and viability on day 14.
- Utilizing morphological assessment and Cell Counting Kit-8 for quantification.
- Investigating adherent cell colonies under different culture conditions.
- Characterizing differentiated cells via immunocytochemistry and induced differentiation assays.
Main Results:
- The optimal plating density for mSKP culture was determined to be 2.5 × 105 to 5 × 105 cells/mL.
- Optimal plating density significantly enhanced the proliferation of adherent cell colonies, improving mSKP growth and yield.
- Adherent cell colonies were found to differentiate into mSKPs or dermal mesenchymal stem cells depending on culture conditions.
- mSKP spheres were confirmed to originate from these adherent cell colonies.
Conclusions:
- Optimal plating density is a critical factor for maximizing mouse SKP growth and yield.
- Adherent cell colonies serve as a source for mSKP spheres and possess multipotent differentiation capacity.
- This study provides a foundation for standardized and efficient mSKP culture protocols.
