Encapsulation of murine hematopoietic stem and progenitor cells in a thiol-crosslinked maleimide-functionalized gelatin hydrogel.
Encapsulation of murine hematopoietic stem and progenitor cells in a thiol-crosslinked maleimide-functionalized gelatin hydrogel.
复制标题
用巯基交联马来酰亚胺功能化明胶水凝胶包封小鼠造血干细胞和祖细胞。
DOI：
10.1016/j.actbio.2021.06.028
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发表时间：
2021-09-01
影响因子：
9.7
通讯作者：
Harley BAC
中科院分区：
文献类型：
--
作者：
Gilchrist AE;Serrano JF;Ngo MT;Hrnjak Z;Kim S;Harley BAC
生物材料平台是干细胞生物制造方案的组成部分。干细胞生态位微环境的生物物理、生物化学和细胞信号在调节干细胞命运决定中起着重要作用。生物材料内干细胞的三维（3D）培养提供了通过分泌的生物分子呈现生物物理和生物化学刺激（例如细胞-基质相互作用和细胞-细胞相互作用）的途径。在此，我们描述了一种马来酰亚胺官能化明胶（GelMAL）水凝胶，其可以通过硫醇-迈克尔加成点击反应交联，用于包封敏感的干细胞群体。沿着明胶骨架的马来酰亚胺官能单元沿着能够通过添加二硫醇交联剂而不需要外部刺激（例如，UV光、光引发剂），从而减少反应性氧化物物质的产生。此外，交联剂选择的多功能性使得能够容易地插入含硫醇的生物活性或生物惰性基序。将造血干细胞（HSC）和间充质干细胞（MSC）包封在GelMAL中，其机械性能被调整以模拟体内骨髓生态位。我们报告插入的可切割的肽交联剂，可以降解的蛋白水解作用的SortaseA，一种非线性的酶。值得注意的是，分选酶A暴露保留了干细胞表面标志物，这是免疫表型分析中使用的造血活性的基本度量。这种新的GelMAL系统能够产生具有可调生物物理特性、内在细胞相互作用基序和正交添加生物活性交联的人工干细胞壁龛。
Biomaterial platforms are an integral part of stem cell biomanufacturing protocols. The collective biophysical, biochemical, and cellular cues of the stem cell niche microenvironment play an important role in regulating stem cell fate decisions. Three-dimensional (3D) culture of stem cells within biomaterials provides a route to present biophysical and biochemical stimuli such as cell-matrix interactions and cell-cell interactions via secreted biomolecules. Herein, we describe a maleimide-functionalized gelatin (GelMAL) hydrogel that can be crosslinked via thiol-Michael addition click reaction for the encapsulation of sensitive stem cell populations. The maleimide functional units along the gelatin backbone enables gelation via the addition of a dithiol crosslinker without requiring external stimuli (e.g., UV light, photoinitiator), thereby reducing reactive oxide species generation. Additionally, the versatility of crosslinker selection enables easy insertion of thiol-containing bioactive or bioinert motifs. Hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) were encapsulated in GelMAL, with mechanical properties tuned to mimic the in vivo bone marrow niche. We report the insertion of a cleavable peptide crosslinker that can be degraded by the proteolytic action of SortaseA, a mammalian-inert enzyme. Notably, SortaseA exposure preserves stem cell surface markers, which are an essential metric of hematopoietic activity used in immunophenotyping. This novel GelMAL system enables a route to produce artificial stem cell niches with tunable biophysical properties, intrinsic cell-interaction motifs, and orthogonal addition of bioactive crosslinks.
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影响因子：
6.2
作者：
Cambria E;Renggli K;Ahrens CC;Cook CD;Kroll C;Krueger AT;Imperiali B;Griffith LG
通讯作者：
Griffith LG
影响因子：
2.1
作者：
Ghasemi A;Zahediasl S
通讯作者：
Zahediasl S
影响因子：
2.5
作者：
Gilchrist, Aidan E.;Harley, Brendan A. C.
通讯作者：
Harley, Brendan A. C.
影响因子：
14.8
作者：
Cruz-Acuña R;Quirós M;Huang S;Siuda D;Spence JR;Nusrat A;García AJ
通讯作者：
García AJ
影响因子：
3.8
作者：
Cuchiara, Maude L.;Coskun, Sueleyman;West, Jennifer L.
通讯作者：
West, Jennifer L.