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B6-Dct-H2BGFP bitransgenic mice: A standardized mouse model for in vivo characterization of melanocyte development and stem cell differentiation
Pigment Cell & Melanoma Research ( IF 4.3 ) Pub Date : 2021-02-05 , DOI: 10.1111/pcmr.12959
Bishal Tandukar 1 , Emmanual Kalapurakal 1 , Thomas J Hornyak 1, 2, 3
Affiliation  

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 inducible Dct-H2BGFP mice into the C57BL/6J background (B6-Dct-H2BGFP). We compared the expression pattern of B6-Dct-H2BGFP to that of Dct-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.

中文翻译:

B6-Dct-H2BGFP 双转基因小鼠:用于体内表征黑素细胞发育和干细胞分化的标准化小鼠模型

黑素细胞干细胞 (McSC) 是毛囊 (HF) 干细胞系统的关键组成部分,可在连续的 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,我们将诱导型Dct -H2BGFP小鼠回交到 C57BL/6J 背景 ( B6- Dct -H2BGFP ) 中。我们在先前报道的混合遗传背景上比较了 B6- Dct -H2BGFPDct -H2BGFP 小鼠的表达模式。表征B6-Dct-H2BGFP 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,而且还证实了强力霉素对 GFP 表达的调节。此外,通过荧光激活细胞分选 (FACS) 分离的 McSC 亚群的离体培养表明,凸起/CD34+ McSC 倾向于通过表达非黑素细胞、神经嵴谱系标记物(包括胶质细胞)(Gfap 和 CNPase,73 ± 1 % 和 77 ± 2%,分别)、神经元(Tuj1 26 ± 5%)和平滑肌(α-Sma,31 ± 9%)。相比之下,CD34-/次级毛胚 (SHG) McSCs 分化为色素黑色素细胞,黑色素标记物 Tyr (71 ± 1%)、Tyrp1 (68 ± 4%) 和 Mitf (75 ± 7%) 的表达更高。这些结果确立了B6-Dct的效用-H2BGFP 双转基因小鼠,用于未来需要明确遗传背景的黑素细胞的体内研究。
更新日期:2021-02-05
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