CN112481200A - 一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法 - Google Patents
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Abstract
本发明涉及一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，包括以下步骤：（1）构建慢病毒干扰载体G9a‑shRNA，将G9a‑shRNA转染至大鼠骨髓间充质干细胞；（2）通过5‑氮杂胞苷诱导转染干扰载体后的骨髓间充质干细胞，在第21天的时候流式细胞检测肌钙蛋白阳性细胞的阳性率。通过本发明，在5‑氮杂胞苷诱导骨髓间充质干细胞分化为心肌细胞过程中，敲低H3K9me2甲基化酶G9a，观察其最终诱导效率，以期为骨髓间充质干细胞分化心肌细胞过程中寻找一种方便、快速的途径。采用FuGENE（Promega）脂质体介导G9a‑shRNA慢病毒重组质粒转染大鼠骨髓间充质干细胞，以减低H3K9me2的表达的方法，从而增加诱导效率，验证结果合理准确，实验过程严密周谨。
Description
技术领域
本发明涉及一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，属于干细胞分化培养领域。
背景技术
生物起搏器为治疗缓慢型心律失常提供了一种新的选择，骨髓间充质干细胞是心脏修复和再生细胞治疗的主要来源之一，因为这些细胞很容易获得，而且对任何不良免疫反应几乎没有固有的免疫原性。但只有非常少的部分间充质干细胞在分化过程中表现为心肌细胞特异性蛋白阳性，这明显限制了其对心脏修复的潜在临床应用。
发明内容
本发明的目的是针对上述问题，提供一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法。
本发明的技术方案是：一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，其特征在于，包括以下步骤：
（1）构建慢病毒干扰载体G9a-shRNA，将G9a-shRNA转染至大鼠骨髓间充质干细胞；
（2）通过5-氮杂胞苷诱导转染干扰载体后的骨髓间充质干细胞，在第21天的时候流式细胞检测肌钙蛋白阳性细胞的阳性率。
步骤（1）中，包括以下步骤：（1-1）根据NCBI提供的G9a的CDS区域序列，设计干扰靶标；合成了上游和下游序列后，退火形成寡核苷酸双链，通过BamHI和EcoRI双酶切连接pGMLV-SC5 RNAi载体得到重组干扰质粒，分别为Sh-G9a；然后将重组干扰质粒进行测序进一步验证；测序成功后，将载体进行慢病毒包；
（1-2）采用FuGENE（Promega）脂质体介导G9a-shRNA慢病毒重组质粒转染大鼠骨髓间充质细胞系；转染前24h，将大鼠骨髓间充质细胞按l×l05/孔接种于24孔板中，用含10%FBS的DMEM培养基培养，DMEM培养基中不含抗生素；观察细胞，待细胞生长状态良好，汇合度达70-80%时，按每孔3μL FuGENE（Promega）+1μg质粒DNA的体系进行转染。
步骤（2）中，包括以下步骤：
处理后的细胞用5-氮杂胞苷诱导；第21天的时候流式细胞检测肌钙蛋白阳性细胞数：细胞在4°C与抗大鼠肌钙蛋白的稀释1:200的一抗孵育过夜，用PBS清洗3遍，然后用1:500的PE-A偶联二抗室温孵育1小时，PBS洗涤5次；流式细胞术检测红色荧光蛋白阳性细胞的百分率。
本发明方法先进科学，通过本发明，提供的一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，在5-氮杂胞苷诱导骨髓间充质干细胞分化为心肌细胞过程中，敲低H3K9me2甲基化酶G9a，观察其最终诱导效率，以期为骨髓间充质干细胞分化心肌细胞过程中寻找一种方便、快速的途径。本方法采用FuGENE（Promega）脂质体介导G9a-shRNA慢病毒重组质粒转染大鼠骨髓间充质干细胞，以减低H3K9me2的表达的方法，从而增加诱导效率，验证结果合理准确，实验过程严密周谨。
本发明的优越性在于：
近年来研究发现，H3K9me2在细胞分化、增殖、重编程等过程密切相关，在心肌细胞成熟过程中，通过调控H3K9me2的表达可以影响心脏发育相关基因表达，保护心脏免受与该基因程序过表达相关的病理性肥大, G9a是继Suv39h1后发现的第二个哺乳动物中甲基化赖氨酸的甲基化修饰酶，主要催化催化常染色质区域的组蛋白H3K9和K27位点。本方法采用慢病毒转染干扰载体敲低H3K9me2甲基化酶G9a用来减低H3K9me2的表达，从而增加5氮杂胞苷诱导大鼠骨髓间充质干细胞分化心肌细胞效率的方法，验证结果合理准确，实验过程严密周谨。验证结果合理准确，实验过程严密周谨。
附图说明
图1为构建G9a慢病毒干扰载体的构建、将G9a干扰载体转染进骨髓间充质干细胞图；
图2为5-氮杂胞苷诱导转染G9a干扰载体的骨髓间充质干细胞图。
具体实施方法
以下实施方式结合附图对本发明做进一步说明。
（1）根据NCBI提供的G9a的CDS区域序列，设计干扰靶标。合成了上游和下游序列后，退火形成寡核苷酸双链，通过BamHI和EcoRI双酶切连接pGMLV-SC5 RNAi载体得到重组干扰质粒，分别为Sh-G9a。然后将重组干扰质粒进行测序进一步验证。测序成功后，将载体进行慢病毒包。
（2）采用FuGENE（Promega）脂质体介导G9a-shRNA慢病毒重组质粒转染大鼠骨髓间充质细胞系。转染前24h，将大鼠骨髓间充质细胞按l×l05/孔接种于24孔板中，用含10%FBS的DMEM培养基(不含抗生素)培养。观察细胞，待细胞生长状态良好，汇合度达70-80%时，按每孔3μL FuGENE（Promega）+1μg质粒DNA的体系进行转染。(图1)。
（3）处理后的细胞用5-氮杂胞苷诱导。第21天的时候流式细胞检测肌钙蛋白阳性细胞数：细胞在4°C与抗大鼠肌钙蛋白的一抗(稀释1:200)孵育过夜，用PBS清洗3遍，然后用PE-A偶联二抗(1:500)室温孵育1小时，PBS洗涤5次。流式细胞术检测红色荧光蛋白阳性细胞的百分率。检测结果如附图2所示。
本实施例操作时使用的相关器材为细胞培养箱、荧光显微镜、载玻片、显微镜、流式细胞仪。
Claims (3)
1.一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，其特征在于，包括以下步骤：
（1）构建慢病毒干扰载体G9a-shRNA，将G9a-shRNA转染至大鼠骨髓间充质干细胞；
（2）通过5-氮杂胞苷诱导转染干扰载体后的骨髓间充质干细胞，在第21天的时候流式细胞检测肌钙蛋白阳性细胞的阳性率。
2.根据权利要求1所述的一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，其特征在于，步骤（1）中，包括以下步骤：（1-1）根据NCBI提供的G9a的CDS区域序列，设计干扰靶标；合成了上游和下游序列后，退火形成寡核苷酸双链，通过BamHI和EcoRI双酶切连接pGMLV-SC5 RNAi载体得到重组干扰质粒，分别为Sh-G9a；然后将重组干扰质粒进行测序进一步验证；测序成功后，将载体进行慢病毒包；
（1-2）采用FuGENE脂质体介导G9a-shRNA慢病毒重组质粒转染大鼠骨髓间充质细胞系；转染前24h，将大鼠骨髓间充质细胞按l×l05/孔接种于24孔板中，用含10%FBS的DMEM培养基培养，DMEM培养基中不含抗生素；观察细胞，待细胞生长状态良好，汇合度达70-80%时，按每孔3μL FuGENE（Promega）+1μg质粒DNA的体系进行转染。
3.根据权利要求1所述的一种提升大鼠骨髓间充质干细胞分化为心肌细胞效率的方法，其特征在于，步骤（2）中，包括以下步骤：
处理后的细胞用5-氮杂胞苷诱导；第21天的时候流式细胞检测肌钙蛋白阳性细胞数：细胞在4°C与抗大鼠肌钙蛋白的稀释1:200的一抗孵育过夜，用PBS清洗3遍，然后用1:500的PE-A偶联二抗室温孵育1小时，PBS洗涤5次；流式细胞术检测红色荧光蛋白阳性细胞的百分率。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276990A (zh) * | 2022-01-13 | 2022-04-05 | 协和华东干细胞基因工程有限公司 | 提高间充质干细胞修复能力的方法及系统 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130244262A1 (en) * | 2010-11-17 | 2013-09-19 | Kyoto University | Cardiomyocyte- and/or cardiac progenitor cell-proliferating agent and method for proliferating cardiomyocytes and/or cardiac progenitor cells |
| CN108977465A (zh) * | 2017-06-02 | 2018-12-11 | 天津医科大学代谢病医院 | 一种靶向沉默Kdm7a基因的shRNA慢病毒载体构建及其应用 |
| CN109022484A (zh) * | 2018-08-07 | 2018-12-18 | 扬州大学 | 一种Nanos2在鸡ESCs向雄性生殖细胞分化过程中功能验证的方法 |
| CN109554351A (zh) * | 2018-12-12 | 2019-04-02 | 山西医科大学 | Rspo1诱导骨髓间充质干细胞向心肌样细胞分化的应用 |
| CN111893091A (zh) * | 2020-08-25 | 2020-11-06 | 扬州大学 | 一种诱导大鼠极小胚胎干细胞分化为心肌细胞的方法 |
-
2020
- 2020-12-25 CN CN202011555174.6A patent/CN112481200A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130244262A1 (en) * | 2010-11-17 | 2013-09-19 | Kyoto University | Cardiomyocyte- and/or cardiac progenitor cell-proliferating agent and method for proliferating cardiomyocytes and/or cardiac progenitor cells |
| CN108977465A (zh) * | 2017-06-02 | 2018-12-11 | 天津医科大学代谢病医院 | 一种靶向沉默Kdm7a基因的shRNA慢病毒载体构建及其应用 |
| CN109022484A (zh) * | 2018-08-07 | 2018-12-18 | 扬州大学 | 一种Nanos2在鸡ESCs向雄性生殖细胞分化过程中功能验证的方法 |
| CN109554351A (zh) * | 2018-12-12 | 2019-04-02 | 山西医科大学 | Rspo1诱导骨髓间充质干细胞向心肌样细胞分化的应用 |
| CN111893091A (zh) * | 2020-08-25 | 2020-11-06 | 扬州大学 | 一种诱导大鼠极小胚胎干细胞分化为心肌细胞的方法 |
Non-Patent Citations (8)
| Title |
|---|
| HO-TAE KIM等: "G9a inhibition promotes neuronal differentiation of human bone marrow mesenchymal stem cells through the transcriptional induction of RE-1 containing neuronal specific genes", 《NEUROCHEMISTRY INTERNATIONAL》 * |
| JINPU YANG等: "Inhibition of G9a Histone Mehtyltransferase Converts Bone Marrow Mesenchymal Stem Cells to Cardiac Competent Progenitors", 《STEM CELLS INTERNATIONAL》 * |
| ROBERTO PAPAIT等: "Histone Methyltransferase G9a Is Required for Cardiomyocyte Homeostasis and Hypertrophy", 《CIRCULATION》 * |
| XIAOLIN SUN等: "H3K9me2 regulates early transcription factors to promote mesenchymal stem-cell differentiation inti cardiomyocytes", 《MOLECULAR MEDICINE REPORTS》 * |
| 买霞等: "5-氮杂胞苷对小鼠骨髓间充质干细胞增殖与分化的影响", 《微量元素与健康研究》 * |
| 卢小媚等: "极小胚胎样干细胞在再生医学中的研究进展", 《医学综述》 * |
| 许皓: "Islet-1促MSCs向心肌样细胞分化中甲基化/乙酰化交互作用机制研究", 《中国博士学位论文全文数据库》 * |
| 阮中宝等: "5-氮杂胞苷诱导人脐带间充质干细胞向心肌样细胞的分化", 《中国组织工程研究与临床修复》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276990A (zh) * | 2022-01-13 | 2022-04-05 | 协和华东干细胞基因工程有限公司 | 提高间充质干细胞修复能力的方法及系统 |
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