毛囊干细胞在微孔阵列装置中均匀聚集的扩增培养,ACS Biomaterials Science & Engineering

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毛囊干细胞在微孔阵列装置中均匀聚集的扩增培养
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2023-02-13 , DOI: 10.1021/acsbiomaterials.2c01141
Sugi Hirano ₁ , Tatsuto Kageyama _{1,

2} , Maki Yamanouchi ₁ , Lei Yan _{1,

2} , Kohei Suzuki _{1,

3} , Katsumi Ebisawa ₄ , Keiichiro Kasai ₅ , Junji Fukuda _{1,

2}

Affiliation

使用毛囊干细胞 (HFSC) 和真皮乳头细胞再生毛发是治疗脱发的一种很有前途的方法。这种方法面临的挑战之一是 HFSC 的定量扩展，同时保持其毛发诱导能力。在这项研究中，HFSC 的扩张是通过形成直径均匀的细胞聚集体实现的，这些细胞聚集体随后被封装在 Matrigel 中。我们设计了一种微孔阵列装置，其中接种小鼠 HFSC，使其形成松散堆积的聚集体 1 小时，然后嵌入 Matrigel 中。定量分析显示，通过这种培养装置，HFSC 数量在 2 周内增加了 20 倍。与典型的扁平基底 Matrigel 悬浮培养细胞相比，设备生长的细胞中毛发干细胞标记物的基因表达显着增加。这些微孔阵列培养的 HFSCs 与新鲜分离的胚胎间充质细胞混合，表明裸鼠皮肤上的毛发再生旺盛。此外，我们检查了这种方法从雄激素性脱发患者中扩增人 HFSCs 的可行性，发现 CD200 的比例与在典型培养皿中或在平坦基质上的 Matrigel 悬浮培养中培养的细胞相比，^{+细胞得到显着改善。}因此，本研究提出的新方法可能对毛发再生医学中的 HFSC 扩增有用。

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Expansion Culture of Hair Follicle Stem Cells through Uniform Aggregation in Microwell Array Devices

Hair regeneration using hair follicle stem cells (HFSCs) and dermal papilla cells is a promising approach for the treatment of alopecia. One of the challenges faced in this approach is the quantitative expansion of HFSCs while maintaining their hair induction capacity. In this study, HFSC expansion was achieved through the formation of uniform-diameter cell aggregates that were subsequently encapsulated in Matrigel. We designed a microwell array device, wherein mouse HFSCs were seeded, allowed to form loosely packed aggregates for an hour, and then embedded in Matrigel. Quantitative analysis revealed a 20-fold increase in HFSC number in 2 weeks through this culture device. Gene expression of trichogenic stem cell markers in the device-grown cells showed a significant increase compared with that of typical flat substrate Matrigel suspension culture cells. These microwell array-cultured HFSCs mixed with freshly isolated embryonic mesenchymal cells indicated vigorous hair regeneration on the skin of nude mice. Furthermore, we examined the feasibility of this approach for the expansion of human HFSCs from androgenetic alopecia patients and found that the ratio of CD200⁺ cells was improved significantly in comparison with that of cells cultured in a typical culture dish or in a Matrigel suspension culture on a flat substrate. Therefore, the novel approach proposed in this study may be useful for HFSC expansion in hair regenerative medicine.

更新日期：2023-02-13

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