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小唾液腺干细胞：临床级培养条件下生物学特性的比较研究。

Minor salivary gland stem cells: a comparative study of the biological properties under clinical-grade culture conditions.

机构信息

Department of Oral Medicine/Pathology, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Greece.

Department of Prosthodontics, Tissue Engineering Core Unit, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Greece.

出版信息

Cell Tissue Res. 2023 Aug;393(2):321-342. doi: 10.1007/s00441-023-03789-z. Epub 2023 May 30.

DOI:10.1007/s00441-023-03789-z
PMID:37249709
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10406694/
Abstract

Development of clinical-grade, cell preparations is central to cGMP (good manufacturing practice compliant) conditions. This study aimed to investigate the potential of two serum/xeno-free, cGMP (StemPro, StemMacs) culture media to maintain "stemness" of human minor salivary gland stem cell (mSG-SC) cultures compared to a complete culture medium (CCM). Overall, StemMacs resulted in higher proliferation rates after p.6 compared to the conventional serum-based medium, while StemPro showed substantial delays in cell proliferation after p.9. The mSG-SCs cultures exhibited two distinct cell populations at early passages a mesenchymal subpopulation and an epithelial-like subpopulation. Expression of several markers (CD146, STRO-1, SSEA-4, CD105, CD106, CD34, K 7/8, K14, K18) variably decreased with prolonged passaging (all three media). The percentage of SA-β-gal positive cells was initially higher for StemMacs compared to StemPro/CCM and increased with prolonged passaging in all cases. The telomere fragment length decreased with prolonged passaging in all three media but more pronouncedly for the CCM. Expansion under serum-free conditions caused pronounced upregulation of ALP and BMP-2, with parallel complete elimination of the baseline expressions of LPL (all three media) and ACAN (serum-free media), therefore, showing a preferential shift of the mSG-SCs towards osteogenic phenotypes. Finally, several markers (Nanog, SOX-2, PDX-1, OTX2, GSC, HCG) decreased with prolonged culture, indicating successive loss of "stemness". Based on the findings, it seems that StemPro preserve stemness of the mSG-SCs after prolonged culture. Nevertheless, there is still a vacant role for the ideal development of clinical-grade culture conditions.

摘要

开发符合 cGMP（良好生产规范）标准的临床级细胞制剂是关键。本研究旨在探讨两种无血清/无动物成分的 cGMP（StemPro、StemMacs）培养基与完整培养基（CCM）相比，维持人小唾液腺干细胞（mSG-SC）培养物“干性”的潜力。总体而言，StemMacs 培养基在 p.6 后比传统基于血清的培养基具有更高的增殖率，而 StemPro 培养基在 p.9 后细胞增殖出现明显延迟。mSG-SC 培养物在早期传代时表现出两种不同的细胞群体，一种是间充质亚群，另一种是上皮样亚群。几种标志物（CD146、STRO-1、SSEA-4、CD105、CD106、CD34、K7/8、K14、K18）的表达随传代时间延长而不同程度降低（所有三种培养基）。与 StemPro/CCM 相比，StemMacs 培养基中 SA-β-gal 阳性细胞的比例最初较高，并且在所有情况下随传代时间延长而增加。在所有三种培养基中，端粒片段长度随传代时间延长而缩短，但 CCM 更为明显。无血清条件下的扩增导致 ALP 和 BMP-2 的显著上调，同时平行完全消除了 LPL 的基线表达（所有三种培养基）和 ACAN（无血清培养基），因此，mSG-SCs 向成骨表型的优先转变。最后，几种标志物（Nanog、SOX-2、PDX-1、OTX2、GSC、HCG）随培养时间延长而降低，表明“干性”逐渐丧失。根据这些发现，StemPro 在延长培养后似乎能保持 mSG-SC 的干性。然而，仍需要进一步开发临床级培养条件。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0acb/10406694/67c493734d7a/441_2023_3789_Fig8_HTML.jpg
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