Expression of pluripotency markers in Arbas Cashmere goat hair follicle stem cells

Abstract

In our previous work, we found that the Inner Mongolia Arbas Cashmere goat hair follicle stem cells (gHFSCs) can be successfully differentiated into adipocyte, chondrocyte, and osteocyte lineages. In this study, we further examined the expression of the pluripotency and stemness markers Oct4, Nanog, Sox2, AKP, and TERT in gHFSCs by immunocytochemistry, flow cytometry, real-time PCR, and Western blot. Immunofluorescent staining showed that the gHFSCs were positive for all five markers. Fluorescence-activated cell sorting (FACS) further analyzed the positive expression of Oct4, Nanog, and Sox2 in the gHFSCs. Compared with Arbas Cashmere goat adipose-derived stem cells (gADSCs) at the mRNA expression level, Oct4 was relatively highly expressed in gHFSCs, 41.36 times of the gADSCs, and Nanog was 5.61, AKP was 2.74, and TERT was 2.10 times, respectively (p < 0.01). Western blot indicated that all markers are expressed at the protein level in the gHFSCs. When compared with gADSCs, using α-tubulin as a reference protein, gray intensity analysis showed that the expression of Oct4, Nanog, AKP, and TERT were, respectively, 5.94, 10.78, 1.33, and 1.39 times of gADSCs. Additionally, mRNA and protein expression of Sox2 were detected in the gHFSCs but not in the gADSCs. The protein expression pattern of these markers was consistent with the mRNA results.

Protective Mechanism of Adipose-Derived Stem Cells in Remodelling of the Skin Stem Cell Niche During Photoaging

“…Skin cells would then tend to overproliferate, which may provide an explanation for UVB-induced skin cancer. He et al showed that the expression of Nanog in goat hair follicle stem cells (gHFSCs) was higher than that in goat adipose-derived stem cells (gADSCs), indicating that the stemness and pluripotency of gHFSCs was stronger than that of gADSCs [26]. We suggest that downregulation of Nanog after grafting of ADSCs may reduce the stemness and pluripotency of skin stem cells and promote differentiation in related cell repair injuries.…”

Hair follicle-associated-pluripotent (HAP) stem cells

“…HAP stem cells differentiate to multiple types of cells HAP stem cells differentiate to multiple types of cells: nerve cells including, adipocytes, glial cells, keratinocytes, smooth muscle cells, cardiac muscle cells, melanocytes, osteocytes and chondrocytes. 19,20 Moreover, HAP stem cells can regenerate the epidermis. The differentiation potential of HAP stem cells is greatest in the upper part of the follicle, which can produce large amounts of these stem cells (Fig.…”

“…A number of laboratories have performed gene-expression analysis on the hair-follicle stem cells, identifying stem-cellrelated gene expression, such as nestin, Nanog, OCT4, Sox-2 as well as differentiation markers 19,34 including human hair follicles stem cells. 35 The gene-expression data are complimentary to the immunohistochemistry data describe above.…”

“…There is controversy regarding "pluripotent" vs "multipotent" designations. We have used the term "pluripotent" since numerous cell types and lineages have shown to differentiate from nestin-expressing hair follicle stem cells including adipocytes, smooth muscle cells, melanocytes, neurons, glial cells, epithelial cells, vascular cells, 2,10,17,[34][35][36] osteocytes and chondrocytes, 19,20 and heart muscle cells. [25][26][27] In addition, as briefly mentioned above, the hair follicle cells express transcription factors found in embryonic stem (ES) cells that are considered "pluripotent" transcription factors by some authors.…”

“…These transcription factors included OCT4, Nanog, Sox-2, AKP, and TERT. 19,34,35 It should be noted that pioneering authors in the field have termed hair-follicle stem cells "pluripotent." 17 Other authors may prefer the term "multipotent.…”