Bo-Young Yoo

Application of mesenchymal stem cells derived from bone marrow and umbilical cord in human hair multiplication

BACKGROUND The methods currently used for treating alopecia have some limitations. The drug treatment is so temporary that medication discontinuance may progress depilation immediately. The number of hair transplantation restricts because total transplantable hair number is no increase. To overcome these problems, researchers have attempted the in vitro culturing of hair follicle cells and implanting these cells in the treatment area. OBJECTIVES In the present study, culture-expanded mesenchymal stem cells (MSCs) that do not possess aggregative activity were used to produce self-aggregated cell-aggregated spheroidal dermal papilla like tissues (DPLTs) with the aid of a special culture condition in vitro, and hair bulb structure inductive capacity pertinent to the aggregative activity was then evaluated. Then hair inducing activity of self-aggregated DPLTs employing MSCs was tested in athymic mice. METHODS We isolated and cultivated MSCs from bone marrow and umbilical cord in vitro. After propagated MSCs underwent preconditioning in dermal papilla forming medium (DPFM), then subcultured MSCs formed self-aggregated DPLTs. We compared real human scalp dermal papilla cells (hDPCs) with DPLTs employing DPCs, DPLTs employing hBM-MSCs and DPLTs employing hUC-MSCs. RESULTS Light microscopy and immunohistochemical staining were used to confirm that reconstructed DPLTs generated by this procedure had the size, shape, and expression of protein similar to actual DP. CONCLUSIONS The DPLTs have the same hair bulb structure inductive ability as natural DPLTs in vitro. Transplanted DPLTs can induce new hair follicle in athymic mice. As a result, UC-MSCs and BM-MSCs may be an applicable and novel cell source for the generation of human hair cell therapy.

Improved isolation of outer root sheath cells from human hair follicles and their proliferation behavior under serum-free condition

Hair follicle is a small but very complex and dynamic miniorgan of the human body. It is easy to isolate and culture mesenchymal cells but not epithelial cells of hair follicle. It is necessary for intact and healthy outer root sheath (ORS) cells to be isolated and cultured. In this study we developed an appropriate isolation method to yield 6.4±0.75×104 cells/hair follicle, which is about 9-fold comparing to our previous data. This yield was achieved by modifications such as different kinds of enzyme uses, fragmentation, and mechanical stimuli. Especially we detected that the different kinds of isolation enzyme could affect proliferation of ORS cells during primary culture. In addition, bovine pituitary extract (BPE) was needed for ORS cells to proliferate and to form colonies under serum-free, feeder layer-free culture condition, but type I collagen as a substratum did not have any positive effect. Moreover, ORS cells under BPE-added condition contained stem/progenitor cells expressing β1-integrin. CK19, and CD34. These results can provide useful cell culture information, not only in the study of hair biology but also in the field of tissue engineering and cell therapy for the treatment of alopecia.

Wound-healing effects of human dermal components with gelatin dressing

There have been numerous investigations regarding various types of dressings and artificial dermis of solid form, yet limited research and development on paste types, such as hydrogels with dermal powder, have ensued. In this study, we compared the in vivo wound healing effects of gelatin paste containing dermal powder to a collagen type I/chondroitin 6-sulfate (coll/chondroitin) sponge and gelatin alone, after 48 days post grafting, in a skin wound rat model. In the dermis powder/gelatin paste-treated group, wound area contraction was minimized 50%, while in the gelatin and coll/chondroitin sponge groups, the initial area contracted 83–85% and 79–85%, respectively. Histological analysis revealed the wounds treated with dermal powder/gelatin were associated with many fibroblasts, which infiltrated the wound bed, as well as thick collagen bundles that were arranged in dendritic arrays, resembling normal skin. Furthermore, in contrast to the gelatin- and coll/chondroitin sponge-treated groups, the powder/gelatin paste-treated wounds exhibited an abundance of elastic fibers (Victoria blue staining) and extensive formation of blood vessels around the dermis (CD31 staining). Therefore, the dermis powder/gelatin paste not only renders convenience to users but also has prominent wound-healing effects on full-thickness wounds.

Hair Multiplication with Dermal Papilla like Tissue Containing Human Dermal Papilla Cells

Alopecia is not a critical disease; however it is a disease that can affect the quality of life. Many remedies have been developed to cure alopecia, but only two have been approved by the FDA. Due to the steadily increasing number of young alopecia patients, the need for new therapies for curing alopecia is very high. Recent studies on cell therapy have reported using technique to treat various diseases. We introduce upgraded hair cell therapy which tested hair structure inducing activity with bioartificial dermal papilla tissue. Hair follicles contain two types of stem cells: Outer root sheath cells (ORSCs) derived epithelial cells, and dermal cells (DPCs). In this study, we reconstructed DP-like tissues (DPLTs) using cultured dermal papilla cells (DPCs) from human hair follicles. The DPLTs were produced special media (Dermal Papilla Forming Media: DPFM) conditions in vitro, which can induce epithelial stands from implanted healthy hair without DP. We tested in vivo hair-inducing with a modified hair sandwich model. Two to three weeks DPLT injection into the mouse scalp skin, we observed new hair in the injected site and detected injected human cells from DPLTs and Outer Root Sheath Cells (ORSCs) in the new hair via human Alu-DNA-specific probe. In the future, reconstructed DPLTs may be used in in vitro studies of hair development and the morphogenesis mechanism, as well as in vitro studies of the efficacy and toxicity of drugs for baldness. These tissues will be used as an alternative medicine product for hair transplantation