Department of Chemical Engineering, Barnett Institute of Chemical and Biological Analysis, Northeastern University , Boston, Massachusetts 02115, USA.
Tissue Eng Part C Methods. 2013 Oct;19(10):765-73. doi: 10.1089/ten.TEC.2012.0638. Epub 2013 Mar 18.
Bulge stem cells reside in the lowest permanent portion of hair follicles and are responsible for the renewal of these follicles along with the repair of the epidermis during wound healing. These cells are identified by surface expression of CD34 and the α6-integrin. When CD34 and α6 double-positive cells are isolated and implanted into murine skin, they give rise to epidermis and hair follicle structures. The current gold standard for isolation of these stem cells is fluorescence-activated cell sorting (FACS) based on cell surface markers. Here, we describe an alternative method for CD34 bulge stem cell isolation: a microfluidic platform that captures stem cells based on cell surface markers. This method is relatively fast, requiring 30 min of time from direct introduction of murine skin tissue digestate into a two-stage microfluidic device to one-pass elution of CD34(+) enriched cells with a purity of 55.8% ± 5.1%. The recovered cells remain viable and formed colonies with characteristic morphologies. When grown in culture, enriched cells contain a larger α6(+) population than un-enriched cells.
隆起部干细胞位于毛囊的最低永久部分，负责这些毛囊的更新以及在伤口愈合过程中表皮的修复。这些细胞通过表面表达 CD34 和 α6-整合素来识别。当分离并植入 CD34 和 α6 双阳性细胞到小鼠皮肤中时，它们会产生表皮和毛囊结构。目前分离这些干细胞的金标准是基于细胞表面标志物的荧光激活细胞分选 (FACS)。在这里，我们描述了一种用于 CD34 隆起部干细胞分离的替代方法：一种基于细胞表面标志物的微流控平台。这种方法相对较快，从直接将小鼠皮肤组织消化物引入两阶段微流控装置到一次性洗脱 CD34(+)富集细胞，仅需 30 分钟，纯度为 55.8%±5.1%。回收的细胞保持活力，并形成具有特征形态的集落。在培养中，富集细胞比未富集细胞含有更大的 α6(+)群体。
