Derivation of Functional Smooth Muscle Cells from Multipotent Human Hair Follicle Mesenchymal Stem Cells

Abstract

We investigated the potential of human hair follicle cells for multilineage differentiation and as a source of functional smooth muscle cells (SMCs). We report that human hair follicle stem cells (HFCs) isolated from individual follicles expressed surface markers that are characteristic of mesenchymal stem cells such as CD44, CD49b, CD73, CD90, and CD105 but lacked hematopoietic markers CD45 and CD34. In addition, HFCs differentiated toward adipocytes, chondrocytes, osteoblasts, or SMCs in the appropriate induction medium. Treatment with basic fibroblast growth factor increased proliferation and prevented myogenic differentiation, suggesting that basic fibroblast growth factor can be used to expand the population of undifferentiated HFCs to the large numbers needed for therapeutic applications. SMCs were isolated from HFCs using tissue-specific promoters and flow cytometry sorting. Cylindrical vascular constructs engineered with HF-SMCs showed remarkable contractility in response to receptor and nonreceptor agonists such KCl, endothelin-1, and the thromboxane mimetic, U46619, as well as superior mechanical properties compared to their counterparts with human vascular SMCs. Our results suggest that HF is a rich source of mesenchymal stem cells with great potential for myogenic differentiation providing functional SMCs for tissue regeneration and cell therapies.

Cadherin-11 regulates mesenchymal stem cell differentiation into smooth muscle cells and development of contractile function in vivo

“…To test this hypothesis, we employed MSCs derived from human bone marrow (BM-MSCs) or human hair follicle (HF-MSCs). The latter were derived in our laboratory and were shown to be clonally multipotent, as individual cells could be induced to differentiate into fat, bone, cartilage and SMCs (Bajpai et al, 2012;Liu et al, 2008;Liu et al, 2010). BM-MSCs or HF-MSCs were plated at low density (3610 3 cells/cm 2 ) or high density (30610 3 cells/cm 2 ) in growth medium [DMEM supplemented with 10% MSC-FBS and 1 ng/ml basic fibroblast growth factor (bFGF)] and, 5 days later, the cells were immunostained for the myogenic proteins, a-smooth muscle actin (aSMA, officially known as ACTA2) and calponin-1 (CNN1).…”

“…Compaction of three-dimensional fibrin hydrogels and contractile force measurements were performed as described previously (Han et al, 2010;Liu et al, 2008;Liu et al, 2010). Briefly, contractile force measurements were performed using an isolated tissue bath system, and the isometric contraction was recorded using a PowerLab data acquisition unit and analyzed with Chart5 software (ADInstruments, Colorado Springs, CO).…”

A novel lentivirus for quantitative assessment of gene knockdown in stem cell differentiation

“…BM-MSCs were purchased from Stem Cell Technologies (Vancouver, Canada) and HF-MSCs were isolated as described previously. 47,48 Both cells were cultured in GM constituted of DMEM supplemented with 5% (v/v) fetal bovine serum and 1% (v/v) antibiotic-antimycotic and 1.0 ng ml --1 basic fibroblast growth factor (BD Biosciences, San Jose, CA, USA). The GM for HASMCs (GIBCO) constituted of 231 Medium (GIBCO) that was supplemented with smooth muscle growth supplement (SMGS; GIBCO).…”

Hair Follicle: A Novel Source of Multipotent Stem Cells for Tissue Engineering and Regenerative Medicine

“…1,2 HFs are easily accessible and contain stem cells from diverse developmental origins that continuously self-renew, differentiate, regulate hair growth, and contribute to skin homeostasis. Hair follicle stem cells (HF-SCs) have been shown to be highly proliferative in vitro and multipotent, [3][4][5] thereby enabling engineering of various tissues for organ replacement and regenerative medicine. In addition, genetic engineering of the HF stem cells in vivo has shown promising results, suggesting that treatment of genetic diseases of skin or hair via HF-SC engineering may be feasible.…”