组织工程与重建外科杂志
組織工程與重建外科雜誌
조직공정여중건외과잡지
JOURNAL OF TISSUE ENGINEERING AND RECONSTRUCTIVE SURGERY
2014年
6期
301-304
,共4页
王玲%唐郑雅%付炜%张文杰
王玲%唐鄭雅%付煒%張文傑
왕령%당정아%부위%장문걸
胚胎干细胞%残留%血小板内皮细胞黏附分子-1
胚胎榦細胞%殘留%血小闆內皮細胞黏附分子-1
배태간세포%잔류%혈소판내피세포점부분자-1
Embryonic stem cells%Residual%Platelet endothelial cell adhesion molecule-1
目的：以血小板内皮细胞黏附分子-1（Platelet endothelial cell adhesion molecule-1，PECAM-1）为标志，去除小鼠胚胎干细胞（Embryonic stem cells，ESCs）中残留未分化的细胞，以去除其致瘤性，为ESCs在研究中的安全应用提供思路。方法将小鼠R1胚胎干细胞株在撤去白血病抑制因子的培养基中悬浮培养6 d，体外自发分化形成类胚体，消化打散后，以PECAM-1为标志进行磁珠分选，得到阳性与阴性细胞群体，分别以2×106个/点注射入裸鼠背部皮下，6~8周后观察畸胎瘤形成情况，组织学分析瘤体构成。结果裸鼠背部成瘤结果显示，PECAM-1+细胞群注射8个点中7个成瘤，成瘤率87.5%；而PECAM-1-细胞群注射8个点中1个成瘤，成瘤率12.5%。 PECAM-1+细胞群与PECAM-1-细胞群成瘤率具有统计学差异（P=0.01）。结论应用PECAM-1可去除体外分化过程中的残留未分化ESCs，并去除其致瘤性。
目的：以血小闆內皮細胞黏附分子-1（Platelet endothelial cell adhesion molecule-1，PECAM-1）為標誌，去除小鼠胚胎榦細胞（Embryonic stem cells，ESCs）中殘留未分化的細胞，以去除其緻瘤性，為ESCs在研究中的安全應用提供思路。方法將小鼠R1胚胎榦細胞株在撤去白血病抑製因子的培養基中懸浮培養6 d，體外自髮分化形成類胚體，消化打散後，以PECAM-1為標誌進行磁珠分選，得到暘性與陰性細胞群體，分彆以2×106箇/點註射入裸鼠揹部皮下，6~8週後觀察畸胎瘤形成情況，組織學分析瘤體構成。結果裸鼠揹部成瘤結果顯示，PECAM-1+細胞群註射8箇點中7箇成瘤，成瘤率87.5%；而PECAM-1-細胞群註射8箇點中1箇成瘤，成瘤率12.5%。 PECAM-1+細胞群與PECAM-1-細胞群成瘤率具有統計學差異（P=0.01）。結論應用PECAM-1可去除體外分化過程中的殘留未分化ESCs，併去除其緻瘤性。
목적：이혈소판내피세포점부분자-1（Platelet endothelial cell adhesion molecule-1，PECAM-1）위표지，거제소서배태간세포（Embryonic stem cells，ESCs）중잔류미분화적세포，이거제기치류성，위ESCs재연구중적안전응용제공사로。방법장소서R1배태간세포주재철거백혈병억제인자적배양기중현부배양6 d，체외자발분화형성류배체，소화타산후，이PECAM-1위표지진행자주분선，득도양성여음성세포군체，분별이2×106개/점주사입라서배부피하，6~8주후관찰기태류형성정황，조직학분석류체구성。결과라서배부성류결과현시，PECAM-1+세포군주사8개점중7개성류，성류솔87.5%；이PECAM-1-세포군주사8개점중1개성류，성류솔12.5%。 PECAM-1+세포군여PECAM-1-세포군성류솔구유통계학차이（P=0.01）。결론응용PECAM-1가거제체외분화과정중적잔류미분화ESCs，병거제기치류성。
Objective To prevent teratoma formation by removal of the residual undifferentiated cells from differentiated mouse embryonic stem cells (mESCs) based on PECAM-1 expression, to hopefully promote the safe use of ESCs-based treatment in future. Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies (EBs) in the absence of leukemia inhibitory factor for 6 days. EBs were then digested into single cells and sorted by magnetic activated cell sorting (MACS) based on PECAM-1 expression. Total of 2×106 PECAM-1+and PECAM-1-cells were injected subcutaneously into nude mice respectively. Teratoma formation was measured after 6-8 weeks. Results Seven out of 8 injected points formed teratoma in the PECAM-1+ cells after 8 weeks of injection, with a tumor formation rate of 87.5%. While only 1 out of 8 injected points formed teratoma in the PECAM-1-cells, with a tumor formation rate of 12.5%. A significantly difference was observed between PECAM-1 positive and negative cells (P=0.01). Conclusion PECAM-1 is a specific marker for residual cells, which could be utilized to remove residual undifferentiated ESCs from in vitro differentiated ESCs, and eventually solve the tumorigenicity problem of ESCs.
