A novel technique for nerve regeneration with adult human epithelial progenitor/"stem cells" keratinocyte

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K19077
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 57040:Regenerative dentistry and dental engineering-related
• Research Institution: The Nippon Dental University
• Principal Investigator: Miyazawa Atsuko 日本歯科大学, 生命歯学部, 助教 (00706997)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: 幹細胞 / 再生医療 / ケラチノサイト / 幹細胞様細胞 / 口腔粘膜

Outline of Final Research Achievements

We analyzed the properties of ePUKs (epithekial Pop Up Keratinocytes) by cell size, cell viability, and immunocytofluorescence biomarker staining. The cell size was the smallest and the cell viability was the highest around day4 or 5 after starting cell culture. The growth factors（Epidermal Growth Factor, vascular endothelial growth factor and Fibloblast Growth Factor 2）in the spent media were evaluated by ELISA. In addition, the ePUKs were cultured in the neuronal differentiation media, but the typical structure of neuron could not be detected in the culture. However, our results showed that these ePUKs appear to be progenitor cells, which are small in size, undifferentiated, and have a high proliferative capacity. We believe that ePUKs are suitable for use in medical applications which are like a therapeutic agent for spinal cord injury in the future.

Free Research Field

再生医療

Academic Significance and Societal Importance of the Research Achievements

臨床現場で必要となる質の高い十分量の幹細胞を採取、また生産することは難しい。本研究では口腔粘膜から、より幹細胞に近いヒト口腔粘膜由来のケラチノサイト"epithelial pop-up keratinocytes (ePUKs)" を獲得する事が出来た。今後in vivoでのさらなる評価・検討が必要となるが、再生医療に注目が集まる骨髄や歯髄等の幹細胞に比べて、採取が容易で、かつ一度に大量の幹細胞様ケラチノサイトを生産・培養することができるこのePUKs培養方法は多くの可能性を秘めており、将来の臨床現場でその他の多能性幹細胞に代わる上皮や神経の再生医療の即戦力となるのではないかと考えている。