Feasibility of human hair follicle-derived mesenchymal stem cells/CultiSpher®-G constructs in regenerative medicine

Abstract

The use of human mesenchymal stem cells (hMSCs) in cell therapies has increased the demand for strategies that allow efficient cell scale-up. Preliminary data on the three-dimensional (3D) spinner culture describing the potential use of microcarriers for hMSCs culture scale-up have been reported. We exploited a rich source of autologous stem cells (human hair follicle) and demonstrated the robust in vitro long-term expansion of human hair follicle-derived mesenchymal stem cells (hHF-MSCs) by using CultiSpher(®)-G microcarriers. We analyzed the feasibility of 3D culture by using hHF-MSCs/CultiSpher(®)-G microcarrier constructs for its potential applicability in regenerative medicine by comparatively analyzing the performance of hHF-MSCs adhered to the CultiSpher(®)-G microspheres in 3D spinner culture and those grown on the gelatin-coated plastic dishes (2D culture), using various assays. We showed that the hHF-MSCs seeded at various densities quickly adhered to and proliferated well on the microspheres, thus generating at least hundreds of millions of hHF-MSCs on 1 g of CultiSpher(®)-G within 12 days. This resulted in a cumulative cell expansion of greater than 26-fold. Notably, the maximum and average proliferation rates in 3D culture were significantly greater than that of the 2D culture. However, the hHF-MSCs from both the cultures retained surface marker and nestin expression, proliferation capacity and differentiation potentials toward adipocytes, osteoblasts and smooth muscle cells and showed no significant differences as evidenced by Edu incorporation, cell cycle, colony formation, apoptosis, biochemical quantification and qPCR assays.

Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors

“…Li et al (2015) selected Cultispher‐G due to the microcarrier'smacroporous and biodegradable properties for the culture of human hair follicle‐derived MSCs . Unlike the findings here, they report successful growth of the human follicle‐derived MSCs on Cultispher‐G.…”

“…Unlike the findings here, they report successful growth of the human follicle-derived MSCs on Cultispher-G. However they do not report the growth of the follicle-derived MSCs on other microcarriers [34]. The study we have conducted here would allow for a swift and robust assessment of multiple microcarriers simultaneously and enable researchers and cell therapy developers to identify the optimal microcarrier with respect to proliferation.…”

“…Whilst it is not possible to compare the data directly, it is likely that differences found between these studies to be predominantly related to the source of hMSCs used (umbilical cord and amniotic membrane). Li et al (2015) selected Cultispher-G due to the microcarrier'smacroporous and biodegradable properties for the culture of human hair follicle-derived MSCs [34]. Unlike the findings here, they report successful growth of the human follicle-derived MSCs on Cultispher-G.…”

Location, Isolation, and Identification of Mesenchymal Stem Cells from Adult Human Sweat Glands

“…However, many experiments suggest variations in the differentiation capacity of MSCs from different tissue sources [ 12 ]. There have been studies on human hair follicle MSCs, which act in the metabolism of hair and repair of skin damage and are applied to cell therapies and regenerative medicine [ 13 , 14 ]. SGs and hair follicles are appendages of the skin, and their embryonic development is similar.…”

Functional Melanocytes Obtained From Rabbit Hair Follicle Outer Root Sheath

“…The ORS of hair follicle is a cellular envelope that surrounds the hair shaft [2] and the inner root sheath (IRS). This mini-organ contains a heterogeneous pool of cells, starting from pluripotent neural crest-like stem cells (NCSC-like) [3,4], Lgr6+ stem cells [5], ranging over neural stem cells [6], mesenchymal stem cells [7][8][9][10], keratinocyte stem cells [11], skin cell precursors [12], to the differentiated keratinocytes and melanocytes [13].…”