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Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity
Published on: January 30, 2021
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Assaying Candidate Human Skin Keratinocyte Stem Cells by Determining Their Long-Term Serial Proliferative Output in
Zalitha Pieterse1,2, Pritinder Kaur3,4
1Curtin Medical School, Curtin University, Bentley, WA, Australia. Zalitha.pieterse@curtin.edu.au.
Methods in Molecular Biology (Clifton, N.J.)
|May 4, 2022
Summary
Researchers identified and isolated three epidermal cell fractions from human neonatal foreskin. The keratinocyte stem cells (KSC) fraction demonstrated the highest long-term proliferative capacity in vitro.
Area of Science:
- Dermatology
- Stem Cell Biology
- Cell Biology
Background:
- Stem cells reside in specific niches throughout the body, including the skin epidermis.
- Stem cells are characterized by their significant long-term proliferative capacity in vitro, distinguishing them from their progeny.
- Identifying and isolating specific stem cell populations is crucial for understanding tissue regeneration and development.
Purpose of the Study:
- To develop and describe a technique for isolating distinct epidermal cell fractions from human neonatal foreskin.
- To differentiate and characterize early differentiating (ED), transient amplifying (TA), and keratinocyte stem cells (KSC) based on cell surface markers.
- To evaluate and compare the long-term proliferative output of these isolated epidermal cell fractions.
Main Methods:
- Isolation of three epidermal cell fractions (ED, TA, KSC) from human neonatal foreskin using differential expression of CD49f and CD71 cell surface markers.
- Cultivation of the three isolated cell fractions in parallel using a serial proliferation assay.
- Assessment of long-term proliferative output and proliferative rate over a 2-3 month experimental timeframe.
Main Results:
- The keratinocyte stem cells (KSC) fraction exhibited the highest total cell yield over the 2-3 month assay period.
- The KSC fraction demonstrated a significantly higher proliferative rate compared to the ED and TA fractions (P > 0.05).
- The study successfully differentiated and quantified the proliferative capacities of the three distinct epidermal cell populations.
Conclusions:
- The described technique effectively isolates and distinguishes epidermal cell populations based on stem cell properties.
- Keratinocyte stem cells (KSC) possess superior long-term proliferative capacity compared to early differentiating and transient amplifying cells.
- This assay methodology can be adapted to assess the proliferative potential of other putative stem cell populations using novel markers.
