B6-Dct-H2BGFP bitransgenic mice: A standardized mouse model for in vivo characterization of melanocyte development and stem cell differentiation.
B6-Dct-H2BGFP bitransgenic mice: A standardized mouse model for in vivo characterization of melanocyte development and stem cell differentiation.
复制标题
B6-Dct-H2BGFP 双转基因小鼠：用于黑素细胞发育和干细胞分化体内表征的标准化小鼠模型。
DOI：
10.1111/pcmr.12959
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发表时间：
2021
影响因子：
4.3
通讯作者：
Hornyak,ThomasJ
中科院分区：
文献类型：
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作者：
Tandukar,Bishal;Kalapurakal,Emmanual;Hornyak,ThomasJ
黑素细胞干细胞（McSC）是毛囊（HF）干细胞系统的关键组成部分，在连续的HF周期中再生分化的黑素细胞。为了促进对黑素细胞发育和分化以及McSC的继续研究，我们将可诱导的Dct-H2 BGFP小鼠回交到C57 BL/6 J背景（B6-Dct-H2 BGFP）中。我们将B6-Dct-H2 BGFP的表达模式与先前报道的混合遗传背景下的OfDct-H2 BGFP小鼠进行了比较。为了表征B6-Dct-H2 BGFP小鼠，我们不仅证实了GFP在所有黑素细胞谱系细胞中的表达，而且还证实了多西环素对GFP表达的调节。此外，通过荧光激活细胞分选术（FACS）分离的McSC亚群的离体培养显示，隆突/CD 34 + McSC倾向于分化，表达非黑素细胞、神经嵴谱系标志物，包括神经胶质细胞（GFAP和CNR，分别为73 ± 1%和77 ± 2%）、神经元（Tuj 1，26 ± 5%）和平滑肌（α-Sma，31 ± 9%）。相比之下，CD 34 −/次级毛胚（SHG）McSCs分化为色素性黑素细胞，黑素生成标记物Tyr（71 ± 1%）、Tyrp 1（68 ± 4%）和Mitf（75 ± 7%）的表达更高。这些结果确立了B6-Dct-H2 BGFP双转基因小鼠用于需要确定遗传背景的黑素细胞的未来体内研究的实用性。
Melanocyte stem cells (McSCs) are key components of the hair follicle (HF) stem cell system that regenerate differentiated melanocytes during successive HF cycles. To facilitate continued research on melanocyte development and differentiation and McSCs, we backcrossed inducibleDct‐H2BGFP mice into the C57BL/6J background (B6‐Dct‐H2BGFP). We compared the expression pattern of B6‐Dct‐H2BGFP to that ofDct‐H2BGFP mice on a mixed genetic background reported previously. To characterize B6‐Dct‐H2BGFP mice, we confirmed not only the expression of GFP in all melanocyte lineage cells, but also doxycycline regulation of GFP expression. Furthermore, ex vivo culture of the McSC subsets isolated by fluorescence‐activated cell sorting (FACS) showed the propensity of bulge/CD34+ McSCs to differentiate with expression of non‐melanocytic, neural crest lineage markers including glia (Gfap and CNPase, 73 ± 1% and 77 ± 2%, respectively), neurons (Tuj1 26 ± 5%), and smooth muscle (α‐Sma, 31 ± 9%). In contrast, CD34−/secondary hair germ (SHG) McSCs differentiated into pigmented melanocytes, with higher expression of melanogenic markers Tyr (71 ± 1%), Tyrp1 (68 ± 4%), and Mitf (75 ± 7%). These results establish the utility of B6‐Dct‐H2BGFP bitransgenic mice for future in vivo studies of melanocytes requiring a defined genetic background.