迄今為止，臍帶血來源的造血幹細胞體外擴增培養技術已趨成熟，克服了幹細胞在臨床移植上數量不足的問題。然而，至今尚不清楚在不同體外培養系統下產生的這些造血幹細胞基因組功能差異。本研究的目的為使用有血清及無血清兩種不同體外擴增培養方式的造血幹細胞，分析兩者有差異表現的基因，探討分子功能層面上的關係。實驗首先將體外擴增培養的造血幹細胞進行RNA提取，接著進行微陣列基因表達晶片分析，篩選出在有血清及無血清培養系統下有差異表達的基因，總共有839個基因，再針對這些基因進行功能描述分析，結果顯示大部分的基因為細胞激素及趨化因子，富集在TNF信號通路，調控血清擴增培養的造血幹細胞發炎、免疫反應。此外，我們進一步使用qPCR分析了調控造血幹細胞功能重要的基因CCL2（C-C motif chemokine ligand 2）、TNF（tumor necrosis factor）和FOS（FBJ murine osteosarcoma viral oncogene homolog）。實驗數據顯示使用體外擴增血清培養系統的造血幹細胞將誘導炎症反應和CD38的高表達，我們認為是胎牛血清內含的異種物質可能刺激造血幹細胞在體外擴增的過程中引起發炎表徵並誘導癌症標誌物CD38表達。本研究提供了造血幹細胞在不同體外擴增培養環境下的廣泛轉錄譜，對造血幹細胞的分子調控網路提供新的見解。;To date, different experimental strategies have been developed for the ex vivo expansion of human hematopoietic stem (HSCs) in clinical application. However, it is still unclear to what difference in genomic function in HSCs expansion under different culture systems. In this study, we compared gene-expression profile of ex vivo expanded serum (10% FBS, fetal bovine serum) and serum-free culture systems, and then analyze molecular function of differentially expressed gene using microarray chips. We identified 839 differentially expressed genes between two culture systems. These genes were enriched in TNF regulated inflammatory pathway in FBS culture system. In addition, mRNA expression of CCL2 (C-C motif chemokine ligand 2) 、 TNF (tumor necrosis factor) and FOS (FBJ murine osteosarcoma viral oncogene homolog) was validated by RT-qPCR. Our data revealed that ex vivo expansion of HSCs using FBS culture system would induce the inflammation response and high expression of CD38. It indicated that FBS culture system might cause inflammation pathway and induce the cancer marker CD38 expression during ex vivo expansion of HSCs. This study provided the transcriptional profile, and new insights into the genomic functionality of HSCs under different expanded cultures.
