Novel In Vitro Culture Condition Improves the Stemness of Human Dermal Stem/Progenitor Cells

Abstract

Cell therapy using adult stem cells has emerged as a potentially new approach for the treatment of various diseases. Therefore, it is an essential procedure to maintain the stemness of adult stem cells for clinical treatment. We previously reported that human dermal stem/progenitor cells (hDSPCs) can be enriched using collagen type IV. However, hDSPCs gradually lose their stem cell properties as in vitro passages continue. In the present study, we developed optimized in vitro culture condition to improve the stemness of these hDSPCs. To evaluate whether the stemness of hDSPCs is well sustained in various culture conditions, we measured the expression levels of SOX2, NANOG, and S100B, which are well-known representative dermal progenitor markers. We observed that hDSPCs grown in three-dimensional (3D) culture condition had higher expression levels of those markers compared with hDSPCs grown in two-dimensional (2D) culture condition. Under the 3D culture condition, we further demonstrated that a high glucose (4.5 g/L) concentration enhanced the expression levels of the dermal progenitor markers, whereas O 2 concentration did not affect. We also found that skin-derived precursor (SKP) culture medium was the most effective, among various culture media, in increasing the dermal progenitor marker expression. We finally demonstrated that this optimized culture condition enhanced the expression level of human telomerase reverse transcriptase (hTERT), the proliferation, and the multipotency of hDSPCs, an important characteristic of stem cells. Taken together, these results suggested that this novel in vitro culture condition improves the stemness of hDSPCs.

Hair growth-promoting effect of human dermal stem/progenitor cell-derived conditioned medium

“…NHDFs were plated onto collagen type IV-coated dishes and then sorted on the basis of their ability to adhere to collagen type IV within 5 min (hDSPCs) or within 12 hr (non hDSPCs) at 37°C [11,19,20].…”

“…Compared to normal human dermal fibroblasts (NHDFs), hDSPCs have relatively high proliferative properties and show higher expression levels of dermal stem cell markers such as SOX2 and S100β. They can also differentiate into mesodermal cell types (adipocytes, chondrocytes, and osteoblasts) and neuronal lineage under specific culture conditions [19,20]. Furthermore, the CM of hDSPCs can recover the expression levels of NHDFs specific markers such as collagen, MMP1 and TIMP1 in UVA-irradiated NHDFs.…”

Ciprofloxacin Improves the Stemness of Human Dermal Papilla Cells

“…DPCs have been intensively investigated to possess many advantages for cell therapy approaches. However, many studies also demonstrated the loss of their stemness and inductive activity during the in vitro passages [ 4 , 6 – 8 ].…”

Stemness Characteristics of Human Corneal Endothelial Cells Cultured in Various Media

“…It has been reported that the proliferation and differentiation of stem cells are controlled depending on the media. 14,33,34 Human corneal endothelial cells have been reported not to proliferate in vivo, 2,3 although HCECs proliferate in vitro. Therefore, these results suggest that medium B and medium BE could provide a similar environment to that found in vivo for HCECs.…”