ROCK inhibitor Y-27632 maintains the proliferation of confluent human mesenchymal stem cells ROCK阻害剤Y-27632はコンフルエントなヒト間葉系幹細胞の増殖を維持する

博士論文

収集根拠 : 博士論文（自動収集）

資料形態 : テキストデータ

コレクション : 国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文

Background and Objective: The transplantation of cell sheets of mesenchymal stem cells (MSCs) is expected to be the next generation of periodontal regenerative therapy. An adequate method of multilayering MSCs has yet to be established. When cell sheets proliferate, they usually contract and detach from culture dishes and then the proliferation of cells in the contracted areas is arrested. ROCK-mediated contractile force causes cell contraction. Although multilayer formation medium (MFM) stimulated the proliferation of growth-arrested confluent MSCs, MSCs detached from the culture dish. Therefore, we investigated the effects of ROCK inhibitor Y-27632 on the proliferation and detachment of confluent MSCs, and examined the ability of cells to differentiate within the cell sheets. Material and Methods: Confluent MSCs were cultured in MFM containing transforming growth factor-β1, ascorbic acid and fetal bovine serum either with or without Y-27632. Cell proliferation was examined by bromodeoxyuridine incorporation assays and total DNA measurement. Sheet contractions were examined by light microscopy and stereomicroscopy. Multilayer formations and focal adhesion assembly were observed with confocal microscopy. Characteristic of cells were examined by flow cytometric analysis. Osteoblast lineage differentiation was observed with alkaline phosphatase and alizarin red S staining. Adipocyte lineage differentiation was observed with oil red O staining. Results: The addition of Y-27632 to MFM prevented the cell sheets from detaching and did not inhibit MSC growth. The cell numbers cultured with MFM/Y-27632 were significantly higher than that obtained with MFM-only on day 4. Cell sheets detached from the culture dish on day 4, and the number of bromodeoxyuridine-positive cells in the detached area decreased. Cells in the cell sheets had similar characteristics to primary MSCs, and differentiated into osteoblast and adipocyte lineages. Conclusion: Y-27632 both prevented the MSC sheets from detaching and maintained the multilayered proliferation of confluent MSCs by MFM, and then cells in the sheets had differentiation potency.

長崎大学学位論文 学位記番号:博(医歯薬)甲第609号 学位授与年月日:平成25年8月7日

Author: Kouhei Nakamura, Atsutoshi Yoshimura, Takashi Kaneko, Kayo Sato, Yoshitaka Hara

Citation: Journal of Periodontal Research, 49(3), pp. 363-370; 2014

Nagasaki University (長崎大学), 博士(歯学) (2013-08-07)

http://hdl.handle.net/10069/34119

長崎大学学位論文 [学位記番号]博（医歯薬）甲第609号 [学位授与年月日]平成25年8月7日

(2013-08-07)