Types of cell culture inserts affect cell crosstalk between co-cultured macrophages and adipocytes
Types of cell culture inserts affect cell crosstalk between co-cultured macrophages and adipocytes
复制标题
DOI：
10.1016/j.bbrc.2023.03.068
复制
发表时间：
2023-04-01
影响因子：
3.1
通讯作者：
Nakahara，Taka
中科院分区：
文献类型：
--
作者：
Xiao，Li;Mochizuki，Mai;Nakahara，Taka
细胞培养插入提供了一个体内样的微环境来研究共培养细胞之间的细胞-细胞相互作用。然而，目前还不清楚插入类型是否会影响细胞串扰。在这里，我们开发了一种环保的细胞培养插入物，XL-insert，它可以减少塑料废物，降低成本。在共培养THP-1巨噬细胞和OP9脂肪细胞中，我们比较了XL插入物与两种类型的商业一次性培养插入物，带胶原膜的ken®插入物（colo -inserts）和带塑料膜的Falcon®插入物（PET-inserts）的细胞-细胞相互作用。扫描电镜、免疫分析和成像分析表明，在三种插入物中，xml -inserts允许共培养巨噬细胞和脂肪细胞的细胞因子自由扩散，并为细胞-细胞相互作用提供了较好的体内样微环境。pet插入物显示出细胞间通讯的局限性，因为膜上的一些孔被胞体阻塞，导致细胞因子通过的渗透性大大降低。Col-inserts阻断了大尺寸细胞因子，但允许小尺寸分子渗透，从而改善了OP9脂肪细胞的脂质积累和脂联素分泌。综上所述，我们的数据表明，膜类型和膜孔径对共培养细胞间串扰的影响非常不同。如果改变插入物，以前的一些共培养研究可能会有不同的结果。
Cell culture inserts offer an in vivo-like microenvironment to investigate cell-cell interactions between co-cultivated cells. However, it is unclear if types of inserts affect cell crosstalk. Here, we developed an environment-friendly cell culture insert, XL-insert, which can reduce plastic waste with lower cost. We compared XL insert with two types of commercial disposable culture inserts, Koken® insert with atelocollagen membrane (Col-inserts) and Falcon® inserts with plastic membrane (PET-inserts) on cell-cell interactions in co-cultivated THP-1 macrophages and OP9 adipocytes. Scanning electron microscope, immunoassay and imaging analysis showed that among three types of inserts, XL-inserts allowed cytokines from co-cultivated macrophages and adipocytes to diffuse freely and offered preferable in vivo-like microenvironment for cell-cell interactions. PET-inserts showed limitations for intercellular communication due to some pores being blocked by somas on the membrane that caused much lower permeability for cytokines passing through. Col-inserts blocked large sized cytokines but allowed small sized molecules to permeate resulting in improved lipid accumulation and adiponectin secretion in OP9 adipocytes. Taken together, our data demonstrated that membrane type and pore size on the membrane affect the cross-talk between co-cultivated cells very differently. Some previous co-culture studies might have different results if the inserts were changed.