人牙髓间充质干细胞（hDPSCs）由于具有高增殖能力、多向分化潜力、免疫调节能力、种族问题少和非侵入性获取等优点，被认为是细胞临床治疗的良好细胞来源。然而，传统上hDPSCs是在含有胎牛血清（FBS）的培养基中分离和扩增的，由于安全问题（病毒传播和过敏），这给临床应用带来了障碍。尽管许多研究努力筛选合适的培养基，但迄今为止结果并不乐观。因此，大规模细胞生产迫切需要符合良好生产规范（GMP）标准的培养系统。本研究旨在寻找合适的培养条件来生产临床级hDPSCs，以满足临床细胞治疗的要求，并进一步促进hDPSCs在组织再生或疾病治疗中的应用。我们从在两种不同介质中扩增的九颗正畸牙齿中获得hDPSCs ：符合 GMP 的异种无血清培养基 (AMMS) 和含血清培养基 (SCM)。系统比较了两种培养基的形态、增殖、标志物表达、分化、干细胞性、衰老和细胞因子分泌等细胞特性。在两种培养基中培养的hDPSCs均表现出间充质干细胞（MSCs）的典型特征。然而，我们发现在AMMS中原代培养中形成了更多的细胞集落，并且hDPSC在AMMS中传代培养期间表现出更高的增殖能力、分化潜力和更好的干性维持。hDPSC在分离和扩增时可以改善细胞特性AMMS 中，这可能为大规模细胞制造提供良好的候选培养基。版权所有 © 2023，国际抗癌研究所（George J. Delinasios 博士），保留所有权利。

Human dental pulp mesenchymal stem cells (hDPSCs) are considered to be a good cell source for cell-based clinical therapy, due to the advantages of high proliferation capacity, multilineage differentiation potential, immune regulation abilities, less ethnic concerns and non-invasive access. However, hDPSCs were traditionally isolated and expanded in medium containing fetal bovine serum (FBS), which is a barrier for clinical application due to the safety issues (virus transmission and allergy). Although many studies make efforts to screen out a suitable culture medium, the results are not promising so far. Therefore, a standard good manufacturing practice (GMP) compliant culture system is urgently required for the large-scale cell production. This study aimed to find suitable culture conditions for producing clinical grade hDPSCs to meet the requirements for clinical cell-based therapy and further to promote the application of hDPSCs into tissue regeneration or disease cure.We derived hDPSCs from nine orthodontic teeth expanded in two different media: a GMP compliant and xenogeneic serum-free medium (AMMS) and a serum containing medium (SCM). Cell propterties including morphology, proliferation, marker expression, differentiation, stemness, senescence and cytokine secretion between these two media were systematically compared.hDPSCs cultured in both media exhibited the typical characteristics of mesenchymal stem cells (MSCs). However, we found that more cell colonies formed in the primary culture in AMMS, and the hDPSCs displayed higher proliferation capacity, differentiation potential and better stemness maintenance during sub-culturing in AMMS.Cell properties of hDPSCs could be improved when they were isolated and expanded in AMMS, which might provide a good candidate of culture medium for large-scale cell manufacturing.Copyright © 2023, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.