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Updated: May 12, 2026
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Isolation of Papillary and Reticular Fibroblasts from Human Skin by Fluorescence-activated Cell Sorting
Published on: May 7, 2019
Improved methods for reprogramming human dermal fibroblasts using fluorescence activated cell sorting.
David J Kahler1, Faizzan S Ahmad, Anita Ritz
1The New York Stem Cell Foundation, New York, New York, United States of America. dkahler@nyscf.org
Plos One
|April 5, 2013
Summary
This study introduces fluorescence-activated cell sorting (FACS) to efficiently isolate induced pluripotent stem cells (iPSCs) from fibroblasts. This method accelerates the generation of high-quality iPSC lines, reducing costs and improving experimental reliability.
Area of Science:
- Stem Cell Biology
- Cellular Reprogramming
- Biotechnology
Background:
- Generating induced pluripotent stem cell (iPSC) lines from human dermal fibroblasts typically involves lengthy and costly viral infection protocols.
- Identifying fully reprogrammed iPSC clones early is challenging due to mixed cell populations, leading to increased maintenance and unreliable experimental outcomes.
Purpose of the Study:
- To develop an improved, faster, and more cost-effective method for deriving high-quality iPSC lines.
- To utilize fluorescence-activated cell sorting (FACS) for prospective isolation of fully reprogrammed iPSCs.
Main Methods:
- Employing FACS to isolate single cells expressing specific surface markers (CD13NEG SSEA4POS Tra-1-60POS) 7-10 days post-viral infection.
- Using both integrating (retroviral) and non-integrating (Sendai virus) reprogramming vectors for iPSC generation.
- Deriving and characterizing 228 iPSC lines from 76 unique patient samples.
Main Results:
- FACS successfully isolated fully reprogrammed iPSCs while depleting parental and partially reprogrammed fibroblasts.
- This method significantly reduced the time and reagents needed for iPSC derivation without small molecule cocktails.
- FACS-derived iPSC lines exhibited pluripotency markers and differentiation potential in vitro and in vivo.
Conclusions:
- FACS offers a robust and efficient strategy for high-throughput iPSC generation.
- This technique streamlines the derivation process, enhancing the quality and reliability of iPSC lines for research.
- The method is applicable to various reprogramming vectors and diverse patient sample sources.
