Gonzalez Hemil, Narasipura Srinivas D, Shull Tanner, Shetty Amogh, Teppen Tara L, Naqib Ankur, Al-Harthi Lena
Department of Internal Medicine, Division of Infectious Diseases, Rush University Medical Center, Chicago, IL 60612, USA.
Department of Microbial Pathogens and Immunity, Rush Medical College, Chicago, IL 60612, USA.
Cells. 2023 Sep 26;12(19):2357. doi: 10.3390/cells12192357.
Human inducible pluripotent stem cell (hiPSC)-derived astrocytes (iAs) are critical to study astrocytes in health and disease. They provide several advantages over human fetal astrocytes in research, which include consistency, availability, disease modeling, customization, and ethical considerations. The generation of iAs is hampered by the requirement of Matrigel matrix coating for survival and proliferation. We provide a protocol demonstrating that human iAs cultured in the absence of Matrigel are viable and proliferative. Further, through a side-by-side comparison of cultures with and without Matrigel, we show significant similarities in astrocyte-specific profiling, including morphology (shape and structure), phenotype (cell-specific markers), genotype (transcriptional expression), metabolic (respiration), and functional aspects (glutamate uptake and cytokine response). In addition, we report that, unlike other CNS cell types, such as neuronal progenitor cells and neurons, iAs can withstand the absence of Matrigel coating. Our study demonstrates that Matrigel is dispensable for the culture of human iPSC-derived astrocytes, facilitating an easy, streamlined, and cost-effective method of generating these cells.
人诱导多能干细胞（hiPSC）来源的星形胶质细胞（iAs）对于研究健康和疾病状态下的星形胶质细胞至关重要。在研究中，它们相对于人胎儿星形胶质细胞具有多个优势，包括一致性、可获得性、疾病建模、定制性以及伦理考量等。Matrigel基质包被是iAs存活和增殖所必需的，这阻碍了iAs的生成。我们提供了一个方案，证明在无Matrigel的情况下培养的人iAs具有活力且能增殖。此外，通过对有和无Matrigel培养物的并列比较，我们展示了星形胶质细胞特异性分析方面的显著相似性，包括形态（形状和结构）、表型（细胞特异性标志物）、基因型（转录表达）、代谢（呼吸）以及功能方面（谷氨酸摄取和细胞因子反应）。另外，我们报告称，与其他中枢神经系统细胞类型（如神经祖细胞和神经元）不同，iAs能够耐受无Matrigel包被的情况。我们的研究表明，Matrigel对于人iPSC来源的星形胶质细胞培养并非必需，这为生成这些细胞提供了一种简便、简化且经济高效的方法。
