Human mesenchymal stem cells from dental pulp (hMSC-DP), including dental pulp stem cells from permanent teeth and exfoliated deciduous teeth, possess unique MSC characteristics such as expression of specific surface molecules and a high proliferation rate. Since hMSC-DP have been applied in numerous clinical studies, it is necessary to establish criteria to evaluate their potency for cell-based therapies. We compared stem cell properties of hMSC-DP at passages 5, 10 and 20 under serum (SE) and serum-free (SF) culture conditions. Cell morphology, proliferation capacity, chromosomal stability, surface phenotypic profiles, differentiation and immunoregulation ability were evaluated. In addition, we assessed surface molecule that regulates hMSC-DP proliferation and immunomodulation. hMSC-DP exhibited a decrease in proliferation rate and differentiation potential, as well as a reduced expression of CD146 when cultured under continuous passage conditions. SF culture conditions failed to alter surface marker expression, chromosome stability or proliferation rate when compared to SE culture. SF-cultured hMSC-DP were able to differentiate into osteogenic, adipogenic and neural cells, and displayed the capacity to regulate immune responses. Notably, the expression level of CD146 showed a positive correlation with proliferation, differentiation, and immunomodulation, suggesting that CD146 can serve as a surface molecule to evaluate the potency of hMSC-DP. Mechanistically, we found that CD146 regulates proliferation and immunomodulation of hMSC-DP through the ERK/p-ERK pathway. This study indicates that SF-cultured hMSC-DP are appropriate for producing clinical-grade cells. CD146 is a functional surface molecule to assess the potency of hMSC-DP.

中文翻译：

---

CD146控制人牙髓临床级间充质干细胞的质量

人牙髓间充质干细胞（hMSC-DP），包括恒牙和脱落乳牙的牙髓干细胞，具有独特的MSC特性，如表达特定的表面分子和高增殖率。由于 hMSC-DP 已应用于众多临床研究，因此有必要建立标准来评估其对基于细胞的治疗的效力。我们比较了血清 (SE) 和无血清 (SF) 培养条件下第 5、10 和 20 代 hMSC-DP 的干细胞特性。评估细胞形态、增殖能力、染色体稳定性、表面表型谱、分化和免疫调节能力。此外，我们评估了调节 hMSC-DP 增殖和免疫调节的表面分子。在连续传代条件下培养时，hMSC-DP 的增殖率和分化潜能降低，CD146 的表达降低。与 SE 培养相比，SF 培养条件未能改变表面标记表达、染色体稳定性或增殖率。SF培养的hMSC-DP能够分化成成骨细胞、脂肪细胞和神经细胞，并显示出调节免疫反应的能力。值得注意的是，CD146 的表达水平与增殖、分化和免疫调节呈正相关，表明 CD146 可以作为表面分子来评估 hMSC-DP 的效力。机制上，我们发现 CD146 通过 ERK/p-ERK 通路调节 hMSC-DP 的增殖和免疫调节。该研究表明，SF 培养的 hMSC-DP 适用于生产临床级细胞。CD146 是一种功能性表面分子，用于评估 hMSC-DP 的效力。