通过条件培养基共培养将人脐带间充质干细胞分化为功能性肠上皮细胞。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-10-18 DOI:10.1016/j.gene.2024.149008

Jiale Hu , Rui Liang , Man Li , Xianglian Zhang , Menglong Li , Huaidong Qu , Zhiqiang Wang

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引用次数: 0

摘要

背景：诱导干细胞分化生成具有吸收功能的肠上皮细胞（IECs），为治疗克罗恩病、溃疡性结肠炎、放射性肠炎和其他难治性肠上皮损伤等疾病提供了巨大的治疗潜力。人脐带间充质干细胞（hUC-MSCs）能够分化为功能性 IEC 样细胞：本研究旨在利用条件培养基共培养法诱导 hUC-MSCs 分化为 IECs：方法：使用从人类 IECs 提取的培养基作为诱导培养基，促进 hUC-MSCs 向 IECs 分化。采用倒置显微镜评估细胞形态，并通过免疫荧光染色分析IEC标记物（包括Villin、CK20、CK8和CK18蛋白）的表达。此外，还通过实时定量 PCR 分析量化了 KRT18 等 IEC 标志物的表达水平。通过蔗糖酶活性测定评估了分化 IEC 分泌蔗糖酶的功能：结果：在诱导的第 14 天，hUC-间充质干细胞表现出与 IECs 相似的形态，并显示出 IEC 标志物的表达，包括 KRT18 基因和 Villin、CK20、CK8 和 CK18 蛋白。蔗糖酶活性测定进一步证实，分化细胞具有显著的蔗糖酶活性：结论：条件培养基共培养法能有效诱导 hUC-MSCs 分化为功能性 IECs。

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Differentiation of human umbilical cord mesenchymal stem cells into functional intestinal epithelial cells via conditioned medium co-culture

Background

The induction of stem cell differentiation to generate intestinal epithelial cells (IECs) with absorptive functions offers significant therapeutic potential for treating conditions such as Crohn’s disease, ulcerative colitis, radiation enteritis, and other refractory intestinal epithelial injuries. Human umbilical cord mesenchymal stem cells (hUC-MSCs) are capable of differentiating into functional IEC-like cells.

Objective

This study aimed to induce the differentiation of hUC-MSCs into IECs using a conditioned medium co-culture method.

Method

A culture medium derived from human IECs was used as the inductive medium to facilitate the differentiation of hUC-MSCs into IECs. The cellular morphology was assessed using inverted microscopy, and the expression of IEC markers, including Villin, CK20, CK8, and CK18 proteins, was analyzed via immunofluorescence staining. Furthermore, the expression levels of IEC markers, such as KRT18, were quantified using real-time quantitative PCR analysis. The functionality of the differentiated IECs in terms of sucrase secretion was assessed through sucrase activity assays.

Results

By the 14th day of induction, hUC-MSCs exhibited a morphology similar to IECs and exhibited the expression of IEC markers, including the KRT18 gene and Villin, CK20, CK8, and CK18 proteins. Sucrase activity assays further confirmed that the differentiated cells demonstrated significant sucrase activity.