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人胚胎干细胞在接枝有聚乙二醇联合纳米段的细胞外基质蛋白衍生肽的水凝胶上培养。

Human embryonic stem cells cultured on hydrogels grafted with extracellular matrix protein-derived peptides with polyethylene glycol joint nanosegments.

机构信息

Department of Botany and Microbiology, King Saud University, Riyadh, Saudi Arabia.

Department of Biotechnology, Bharath Institute of Higher Education and Research, Chennai-73, India.

出版信息

IET Nanobiotechnol. 2022 Dec;16(9):295-304. doi: 10.1049/nbt2.12091. Epub 2022 Oct 6.

Abstract

Human pluripotent stem cells (hPSCs) can be proliferated on completely synthetic materials under xeno-free cultivation conditions using biomaterials grafted with extracellular matrix protein (ECM)-derived peptides. However, cell culture biomaterials grafted with ECM-derived peptides must be prepared using a high concentration of peptide reaction solution (e.g. 1000 μg/ml), whereas the ECM concentration of the ECM-coated surface for hPSC culture is typically 5 μg/ml. We designed a polyethylene glycol (PEG) joint nanosegment (linker) to be used between base cell culture biomaterials and bioactive ECM-derived peptides to enhance the probability of contact between ECM-derived peptides and cell binding receptors of hPSCs. Vitronectin-derived peptides with glycine joint nanosegments (GCGG) were conjugated onto poly (vinyl alcohol-co-itaconic acid) hydrogels via PEG joint nanosegments, and human embryonic stem cells (hESCs) were cultivated on these hydrogels. hESCs could successfully be cultivated on hydrogels while maintaining their pluripotency and differentiation potential to differentiate into cells that are induced from three germ layers in vitro and in vivo, where only a 50 μg/ml ECM-derived peptide concentration was used when the PEG joint nanosegments were introduced into peptides that were grafted onto hydrogel surfaces. The joint nanosegments between bioactive peptides and base cell culture biomaterials were found to contribute to efficient hESC attachment and proliferation.

摘要

人多能干细胞(hPSCs)可在无动物培养条件下于完全合成材料上增殖,此时需将细胞外基质蛋白(ECM)衍生肽接枝到生物材料上。然而,将 ECM 衍生肽接枝到细胞培养生物材料上时必须使用高浓度的肽反应溶液(例如 1000μg/ml),而 hPSC 培养用 ECM 涂层表面的 ECM 浓度通常为 5μg/ml。我们设计了一种聚乙二醇(PEG)连接纳米段(接头),用于基础细胞培养生物材料和生物活性 ECM 衍生肽之间,以提高 ECM 衍生肽与 hPSC 细胞结合受体接触的可能性。通过 PEG 连接纳米段,将带有甘氨酸连接纳米段(GCGG)的纤连蛋白衍生肽接枝到聚乙烯醇-co-衣康酸共聚物水凝胶上,并在这些水凝胶上培养人胚胎干细胞(hESCs)。当将 PEG 连接纳米段引入接枝到水凝胶表面的肽中时,hESCs 可以成功地在水凝胶上培养,同时保持其多能性和分化潜能,分化为体外和体内诱导的三个胚层细胞,此时仅使用 50μg/ml 的 ECM 衍生肽浓度。发现生物活性肽和基础细胞培养生物材料之间的连接纳米段有助于 hESC 的有效附着和增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b4f2/9667744/548df0172fc5/NBT2-16-295-g006.jpg

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