[關(guān)鍵詞]
[摘要]
目的 基于轉(zhuǎn)錄組數(shù)據(jù)探討脂肪組織(AD)、胎盤(pán)絨毛膜(HC)、胎盤(pán)羊膜(HA)和臍帶(UC)來(lái)源的間充質(zhì)干細(xì)胞 (MSCs) 的生物學(xué)異質(zhì)性。方法 從人 AD、HC、HA 和 UC 中分離 MSCs,流式細(xì)胞術(shù)檢測(cè)細(xì)胞表面陽(yáng)性標(biāo)志物(CD73、CD90、CD105)和陰性標(biāo)志物(CD14、CD34、CD45、CD79a、HLA-DR)的表達(dá),改良版茜素紅染色、油紅O 染色、阿利辛藍(lán)染色檢測(cè)細(xì)胞的三系分化能力;Trizol 法提取細(xì)胞總 RNA,用于轉(zhuǎn)錄組測(cè)序,應(yīng)用 GFOLD(1.1.4)進(jìn)行差異基因表達(dá)分析;使用 DAVID 數(shù)據(jù)庫(kù)對(duì)差異表達(dá)的基因進(jìn)行基因本體(GO)功能富集分析。結(jié)果 P2 代的不同來(lái)源的 MSCs,CD73、CD90、CD105 均為陽(yáng)性表達(dá),CD14、CD34、CD45、CD79a、HLA-DR 均為陰性表達(dá),培養(yǎng)的 MSCs 均具有三系分化能力。新生兒來(lái)源的 MSCs(HA、HC 和 UC)相關(guān)性大于成人來(lái)源的 MSCs(AD),在功能富集分析中,與來(lái)源于 AD 的MSCs 相比,HA 和 UC 來(lái)源的 MSCs 表現(xiàn)出更優(yōu)異的增殖能力。來(lái)源于 AD 的 MSCs 有更好的分化潛力以及促進(jìn)血管生成能力,而來(lái)源于 UC 的 MSCs 支持神經(jīng)元的發(fā)育并分泌可以調(diào)節(jié)免疫環(huán)境的趨化因子和抗炎因子。結(jié)論 不同來(lái)源的 MSCs 具有不同的生物學(xué)特征,提示不同來(lái)源的 MSCs 可能具有不同臨床應(yīng)用的最佳選擇。
[Key word]
[Abstract]
Objective To investigate the biological heterogeneity of adipose tissue (AD), placental chorionic membrane (HC), placental amniotic membrane (HA) and umbilical cord (UC) derived mesenchymal stem cells (MSCs) based on transcriptome data. Methods MSCs were isolated from human AD, HC, HA and UC, and the expressions of positive markers (CD73, CD90, CD105) and negative markers (CD14, CD34, CD45, CD79a, HLA-DR) on the cell surface were detected by flow cytometry. Improved alizarin red staining, oil red O staining and alisin blue staining were used to detect the three-line differentiation ability of the cells. Total RNA was extracted by Trizol method for transcriptome sequencing, and differential gene expression was analyzed by GFOLD (1.1.4). The gene ontology (GO) functional enrichment analysis of differentially expressed genes was performed using the DAVID database. Results In P2 generation of MSCs from different sources, CD73, CD90 and CD105 were all positively expressed, while CD14, CD34, CD45, CD79a and HLA-DR were all negatively expressed. The cultured MSCs all had the ability of three-line differentiation. Correlation analysis revealed that MSCs derived from neonatal sources (HA, HC, and UC) were more closely related to each other than those derived from adult sources (AD). In functional enrichment analysis, compared to MSCs from AD, those derived from HA and UC showed superior proliferative abilities. AD-derived MSCs demonstrated better differentiation potential and angiogenesis ability. In contrast, UC-derived MSCs supported neuronal development and secreted chemokines and antiinflammatory factors that regulate the immune environment. Conclusion MSCs derived from different sources possess distinct biological characteristics, suggesting that the optimal choice for clinical applications may vary depending on the source of MSCs.
[中圖分類號(hào)]
R965
[基金項(xiàng)目]
國(guó)家重點(diǎn)研發(fā)計(jì)劃干細(xì)胞及相關(guān)治療產(chǎn)品質(zhì)量控制和非臨床評(píng)價(jià)關(guān)鍵技術(shù)與規(guī)范研究項(xiàng)目(2021YFA1101603);天津市科技計(jì)劃項(xiàng)目細(xì)胞產(chǎn)品開(kāi)發(fā)和臨床應(yīng)用的研究(22ZYJDSY00150);天津市科技計(jì)劃項(xiàng)目細(xì)胞制品的成藥性及轉(zhuǎn)化研究(23ZGCX‐QY00050)