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CD44 is a negative cell surface marker for pluripotent stem cell identification during human fibroblast
Rene H Quintanilla1, Joanna S T Asprer1, Candida Vaz2
1Cell Biology & Stem Cell Sciences, Life Technologies, Carlsbad, California, United States of America.
Plos One
|January 14, 2014
Summary
Identifying fully reprogrammed induced pluripotent stem cells (iPSCs) is crucial. Researchers found that CD44 cell surface marker expression helps distinguish partially reprogrammed cells from fully reprogrammed iPSCs, improving clone selection for research and therapy.
Area of Science:
- Stem cell biology
- Cellular reprogramming
- Biomarker discovery
Background:
- Induced pluripotent stem cells (iPSCs) hold significant promise for regenerative medicine and disease modeling.
- Accurate identification of fully reprogrammed iPSC colonies is essential for successful derivation and application.
- Current methods using morphology and pluripotent markers are insufficient to differentiate partially reprogrammed cells from true iPSCs.
Purpose of the Study:
- To identify reliable markers for distinguishing fully reprogrammed iPSCs from partially reprogrammed cells and parental fibroblasts.
- To improve the efficiency and accuracy of iPSC colony selection.
- To reduce the selection of suboptimal clones for downstream applications.
Main Methods:
- Global transcriptome analysis to identify differential gene expression between partially and fully reprogrammed cells.
- Immunohistochemistry and flow cytometry to validate the expression of candidate markers on human fibroblasts and iPSCs.
- Comparative analysis of colony morphology, pluripotent markers, and the novel marker (CD44) for iPSC identification.
Main Results:
- Global transcriptome analysis identified CD44 as a key differentiator between partially and fully reprogrammed cells.
- CD44 is highly expressed in parental fibroblasts, gradually decreases during reprogramming, and is absent in fully reprogrammed iPSCs.
- Combining CD44 staining (negative) with pluripotent cell markers (positive) enables clear and accurate identification of fully reprogrammed iPSC colonies.
Conclusions:
- CD44 serves as a reliable negative surface marker to distinguish fully reprogrammed iPSCs from partially reprogrammed cells.
- The combination of CD44 and pluripotent markers enhances the accuracy and efficiency of iPSC selection.
- This improved selection strategy minimizes the use of suboptimal clones, advancing iPSC-based research and therapeutic development.
