Multipotent stem cells in dental pulp

소개

dental pulp stem cells 에 대한 내용으로서 odontogenic(Osteogenic), adipogenic, neural differentiation medium 조성과, primary culture dental pulp stem cells 에 관한 내용으로서 dental pulp 에서 stem cells을 분리하는데 많은 도움이 될것이다.

목차

Mulripotent stem cells in dental pulp
table 1.2 Preparation of differentiation media
1.3.4. Primary Culture of DPSCs/SHED
Protocol 1.2 Primary Culture of DPSCs or SHED
1.3.5. Subculture
Protocol 1.3. Subculture of DPSCs

본문내용

1.3.4. Primary Culture of DPSCs/SHED
This isolation protocol is based on the ability of stem cells to adhere to culture dishes and form discrete colony clusters [Friedenstein, 1976; Friedenstein et al., 1970]. To obtain colonies each derived from a single cell (i.e., clones), it is very important to release DPSCs and SHED from their perivascular niche in the pulp tissue [Shi and Gronthos, 2003]. Use sterile scalpel blasé and chop up the pulp into small segments. Enzyme digestion is then required for harvesting putative stem cells from pulp tissue. However, it may be harmful for the stem cells of enzyme digestion lasts for more than 1h. Each researcher should find optimal conditions for the digestion. Usually, it takes 30~60min to fully digest well-minced pulp tissue.
It is recommended that the medium be changed 1day after cell isolation; this minimizes the incidence of contamination. However, when the risk of contamination is high, the medium can be changed 5h after the isolation.
Cells isolated from dental pulp are usually seeded at 1-10 x 103 cells/cm2 for culture. To evaluate their colony formation rate (CFU-F assay), a lower cell density (0.1-1.0 X 103cells/cm2) is recommended in order to distinguish each single colony cluster.

참고자료

· 없음