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Stem Cells Transl Med. 2014 Nov;3(11):1354-62. doi: 10.5966/sctm.2014-0098. Epub 2014 Sep 9.

Microfluidic Isolation of CD34-Positive Skin Cells Enables Regeneration of Hair and Sebaceous Glands In Vivo.

Author information

  • 1Department of Chemical Engineering and Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, Massachusetts, USA; Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, Massachusetts, USA; Grass Center for Bioengineering, The Hebrew University of Jerusalem, Jerusalem, Israel; Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
  • 2Department of Chemical Engineering and Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, Massachusetts, USA; Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, Massachusetts, USA; Grass Center for Bioengineering, The Hebrew University of Jerusalem, Jerusalem, Israel; Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA s.murthy@neu.edu.

Abstract

Skin stem cells resident in the bulge area of hair follicles and at the basal layer of the epidermis are multipotent and able to self-renew when transplanted into full-thickness defects in nude mice. Based on cell surface markers such as CD34 and the α6-integrin, skin stem cells can be extracted from tissue-derived cell suspensions for engraftment using the gold standard cell separation technique of fluorescence-activated cell sorting (FACS). This paper describes an alternative separation method using microfluidic devices coated with degradable antibody-functionalized hydrogels. The microfluidic method allows direct injection of tissue digestate (no preprocessing tagging of cells is needed), is fast (45 minutes from injected sample to purified cells), and scalable. This method is used in this study to isolate CD34-positive (CD34+) cells from murine skin tissue digestate, and the functional capability of these cells is demonstrated by transplantation into nude mice using protocols developed by other groups for FACS-sorted cells. Specifically, the transplantation of microfluidic isolated CD34+ cells along with dermal and epidermal cells was observed to generate significant levels of hair follicles and sebaceous glands consistent with those observed previously with FACS-sorted cells.

KEYWORDS:

CD34-positive stem cells; Microfluidic devices; Nude mice; Skin graft site

PMID:
25205844
PMCID:
PMC4214846
DOI:
10.5966/sctm.2014-0098
[PubMed - indexed for MEDLINE]
Free PMC Article
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