| 张晓晓,张 勇,徐 溪,王高文,张 艳,吕 行,吴 朔.脐带间充质干细胞（HUC-MSCs）质体（Cytoplast）来源CD64过表达纳米囊泡的构建方法研究[J].,2024,(7):1209-1215 |
| 脐带间充质干细胞（HUC-MSCs）质体（Cytoplast）来源CD64过表达纳米囊泡的构建方法研究 |
| The Construction of CD64 (FcγRI) Coated Nanovesicle From the Cytoplast of Human Umbilical Cord Mesenchymal Stem Cells (HUC-MSCs) |
| 投稿时间：2023-12-08 修订日期：2023-12-31 |
| DOI：10.13241/j.cnki.pmb.2024.07.002 |
| 中文关键词: 脐带间充质干细胞（HUC-MSCs） 细胞质体（Cytoplast） 纳米囊泡 CD64（FcγRI） |
| 英文关键词: Human umbilical cord mesenchymal stem cells(HUC-MSCs) Cytoplast Nanovesicle CD64(FcγRI) |
| 基金项目:国家自然科学基金青年项目（81902316） |
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| 中文摘要: |
| 摘要 目的：人脐带间充质干细胞（HUC-MSCs）来源的纳米颗粒可通过多种载药方式用于药物靶向，是近年来医药领域的研究热点。本文应用过表达CD64（FcγRI）的脐带间充质干细胞（HUC-MSCs）细胞质体（Cytoplast）为原料，制备可装载IgG1/IgG3靶向抗体的纳米囊泡，并检测纳米囊泡的入胞效率。方法：应用组织块培养法分离HUC-MSCs；建立pLV-CD64慢病毒包装载体并包装慢病毒，用慢病毒感染HUC-MSCs并筛选稳定表达CD64的细胞株；通过细胞松胞菌素B共孵育/离心法去除HUC-MSCs细胞核获得质体；超声法和梯度挤出法制备直径约200 nm的囊泡；将纳米囊泡与抗EGFR的IgG1抗体共孵育以装载靶向抗体，并观察该纳米囊泡进入A549细胞的入胞效率变化。结果：成功获得稳定表达CD64的HUC-MSCs质体，制备装载EGFR抗体的纳米囊泡（粒径为195±82 nm）并有效提高了该囊泡对靶细胞的入胞效率。结论：成功构建可装载IgG1抗体的纳米微泡，该纳米囊泡可高效进入表达EGFR的A549靶细胞。 |
| 英文摘要: |
| ABSTRACT Objective: The nanovesicle derived from human umbilical cord mesenchymal stem cells (HUC-MSCs) can be used for drug targeting through a variety of methods. The aim of this study is to construct the CD64 (FcγRI) coated nanovesicle from the HUC-MSCs, and to detect the efficiency of antibodies (IgG1/IgG3) loading and cell entry of the nanovesicle. Methods: HUC-MSCs were isolated by tissue-cutting and cell culture method; A pLV-CD64 lentivirus packaging vector was established and packaged in the lentivirus; The lentivirus was transfected into HUC-MSCs, and the cell line that stably express CD64 was selected; The CD64 expressing HUC-MSCs was denuclearizated using the cytochalasin B co-incubation/centrifugation method; The nanovesicle with a diameter of approximately 200 nm was generated by ultrasound and gradient extrusion methods. The nanovesicle was co-incubated with anti-EGFR IgG1 antibodies for antibody loading, and the entry efficiency of the nanovesicle was investigated in A549 cells. Results: The nanovesicle with the diameter of 195±82 nm, was obtained using the cytoplast of HUC-MSCs. The nanovesicle was stably expressing CD64 and successfully loaded with EGFR antibody; The modification effectively improved the entry efficiency of the nanovesicle for target cells. Conclusion: A nanovesicle with the capable of loading IgG1 antibodies was constructed, which could efficiently improve its entry efficiency for target cells. |
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