A novel aldehyde dehydrogenase-3 activator leads to adult salivary stem cell enrichment in vivo.
A novel aldehyde dehydrogenase-3 activator leads to adult salivary stem cell enrichment in vivo.
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DOI：
10.1158/1078-0432.ccr-11-0179
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发表时间：
2011-12-01
期刊：
影响因子：
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通讯作者：
Le QT
中科院分区：
文献类型：
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作者：
Banh A;Xiao N;Cao H;Chen CH;Kuo P;Krakow T;Bavan B;Khong B;Yao M;Ha C;Kaplan MJ;Sirjani D;Jensen K;Kong CS;Mochly-Rosen D;Koong AC;Le QT
评估成人和小鼠下颌下腺（SMG）干细胞中醛脱氢酶（ALDH）的表达，并确定ALDH 3激活对SMG干细胞富集的影响。通过细胞表面标志物（人用CD 34，小鼠用c-Kit）选择成人和小鼠SMG干细胞，并通过流式细胞术表征各种其他干细胞表面标志物，通过定量逆转录酶PCR表征ALDH同工酶表达。球体形成和溴脱氧尿苷（BrdUrd）掺入试验用于选定的细胞，以确认其更新能力和三维（3D）胶原基质培养，以观察分化。为了确定ALDH 3活化是否会增加干细胞产量，将成年小鼠输注新型ALDH 3活化剂（Alda-89）或载体，然后定量c-Kit+/CD 90 + SMG干细胞和BrdUrd+盐球。超过99%的CD 34 + huSMG干细胞对c-Kit、CD 90染色呈阳性，70%与CD 44、Nestin共定位。同样，73.8%的c-Kit+ mSMG干细胞与Sca-1共定位，而80.7%与CD 90共定位。在功能上，这些细胞形成BrdUrd+ salispheres，当在3D胶原中培养时，其分化成腺泡和导管样结构。成人和小鼠SMG干细胞均显示ALDH 3的表达高于其非干细胞，并且这些细胞中的84%具有可测量的ALDH 1活性。与对照组相比，成年小鼠中的Alda-89输注显著增加了c-Kit+/CD 90 + SMG群体和BrdUrd+球形成。这是第一个研究不同的ALDH同工酶在SMG干细胞中的表达特征。ALDH 3的体内激活可以增加SMG干细胞产量，从而为将来的干细胞治疗提供了SMG干细胞富集的新手段。
To assess aldehyde dehydrogenase (ALDH) expression in adult human and murine submandibular gland (SMG) stem cells and to determine the effect of ALDH3 activation in SMG stem cell enrichment. Adult human and murine SMG stem cells were selected by cell surface markers (CD34 for human and c-Kit for mouse) and characterized for various other stem cell surface markers by flow cytometry and ALDH isozymes expression by quantitative reverse transcriptase PCR. Sphere formation and bromodeoxyuridine (BrdUrd) incorporation assays were used on selected cells to confirm their renewal capacity and three-dimensional (3D) collagen matrix culture was applied to observe differentiation. To determine whether ALDH3 activation would increase stem cell yield, adult mice were infused with a novel ALDH3 activator (Alda-89) or with vehicle followed by quantification of c-Kit+/CD90+ SMG stem cells and BrdUrd+ salispheres. More than 99% of CD34+ huSMG stem cells stained positive for c-Kit, CD90 and 70% colocalized with CD44, Nestin. Similarly, 73.8% c-Kit+ mSMG stem cells colocalized with Sca-1, whereas 80.7% with CD90. Functionally, these cells formed BrdUrd+ salispheres, which differentiated into acinar- and ductal-like structures when cultured in 3D collagen. Both adult human and murine SMG stem cells showed higher expression of ALDH3 than in their non–stem cells and 84% of these cells have measurable ALDH1 activity. Alda-89 infusion in adult mice significantly increased c-Kit+/CD90+ SMG population and BrdUrd+ sphere formation compared with control. This is the first study to characterize expression of different ALDH isozymes in SMG stem cells. In vivo activation of ALDH3 can increase SMG stem cell yield, thus providing a novel means for SMG stem cell enrichment for future stem cell therapy.