Novel in vitro culture condition improves the stemness of human dermal stem/progenitor cells.
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.
Bioscience Research Institute, Amorepacific Corporation R&D Center, Yongin, 446-729, Korea.
Joong Hyun
JH
Shim
Tae Ryong
TR
Lee
Dong Wook
DW
Shin
D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000494:pharmacology*; D020405:Dermis / Q000166:cytology*; D005947:Glucose / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology / Q000502:physiology; D000071317:Nanog Homeobox Protein; D010100:Oxygen / Q000378:metabolism; D064568:S100 Calcium Binding Protein beta Subunit / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism
D015415:Biomarkers; D024141:Collagen Type IV; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D064568:S100 Calcium Binding Protein beta Subunit; C579160:S100B protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C509186:TERT protein, human; D019098:Telomerase; D005947:Glucose; D010100:Oxygen