Maintenance of high proliferation and multipotent potential of human hair follicle-derived mesenchymal stem cells by growth factors

Abstract

Cell therapy and cell-based tissue engineering is becoming increasingly important in regenerative medicine. Stem cells that are characterized by self-renewal, high proliferation and multiple differentiation potentials have attracted attention in cell-based regenerative medicine. Maintaining the aforementioned characteristics of stem cells is the first key step in cell-based regenerative medicine. Basic fibroblast growth factor (bFGF) is a well-known growth factor that efficiently maintains the self-renewal, high proliferation and multilineage differentiation potential of stem cells. Whether or not other growth factors, such as acidic fibroblast growth factor (aFGF) and epidermal growth factor (EGF) have similar effects has yet to be fully elucidated. Human hair follicle-derived mesenchymal stem cells (HF-MSCs) were obtained by organ culture. They exhibited surface markers of bone marrow mesenchymal stem cells as shown by positive staining for CD44, CD73, CD90 and CD105, and they also displayed trilineage differentiation potentials into adipocytes, chondrocytes and osteoblasts by cytochemistry and qRT-PCR. Flow cytometry analysis showed that up to 70% of HF-MSCs cultured in the presence of aFGF, bFGF or EGF stayed at the G0/G1 phase. Proliferation analysis showed that both bFGF and EGF at as low as 1 ng/ml and aFGF at above 5 ng/ml levels significantly increased the proliferation of HF-MSCs by cell counting. Consistent with proliferation analysis, immunofluorescence staining showed that more than 95% of HF-MSCs cultured in the presence of aFGF, bFGF and EGF were positively stained for proliferating cell nuclear antigen. HF-MSCs cultured in the presence of aFGF, bFGF or EGF retained marked trilineage differentiation potentials. By contrast, HF-MSCs cultured in the absence of bFGF, aFGF and EGF lost multipotency.

Autologous Micrografts from Scalp Tissue: Trichoscopic and Long-Term Clinical Evaluation in Male and Female Androgenetic Alopecia

“…For ST, in previous research conducted by Gentile et al [9], a procedure was tuned to collect HFSCs with minimal manipulation based on the centrifugation of the scalp's fragments obtained by punch biopsy, without cellular extension or culture. In this procedure, we succeeded in cell counting and identifying CD44+ HF-MSCs and CD200+ HF-ESCs, according to previously reported studies [10,11].…”

“…Micrografts' suspension tests (n � 27) were fixed with 4% paraformaldehyde and characterized for mesenchymal CD44 [10] and epithelial CD200 [11] stem cell markers, according to previously reported studies [10,11]. After cytospin, cells were analyzed by immunocytochemistry with specific primary antibodies (CD200 ab203887, 1 : 100; CD44 sc-9960, 1 : 10).…”

Induction of osteoblastic differentiation of neural crest-derived stem cells from hair follicles

“…Those are known to contain stem cells [42–44], with the dermal papilla (DP) in particular reported to retain stem cell-like properties and the hair follicle bulge area (bulge) to contain adult stem cells [42, 43]. In addition, hair follicle stem cells have been shown to have potential for osteoblast differentiation [45–47], although detailed findings indicating their differentiation potential have not been presented.…”

Antigen‐extracted xenogeneic cancellous bone graft with recombinant human bone morphogenetic protein‐2 enhances bone regeneration in repair of mandibular defect in rabbits

“…Autologous bone graft provides not only a scaffold but also a certain number of osteoblasts, and it has the best osteogenic effect. Therefore, it is considered the gold standard for bone defect repair [1–3]. The autologous cancellous bone is usually taken from the iliac cancellous bone, the distal femur, the greater trochanter, or the proximal tibia [4].…”