CD271磁珠体外诱导人胚胎干细胞分化为成骨细胞的可行性

Osteoblasts derived from human embryonic stem cells by CD271 magnetic beads in vitro

背景:来源于人胚胎干细胞的成骨细胞研究结果引人关注,但是能够产生大量有效的成骨细胞且不含有其他杂细胞的诱导方法仍然没有建立起来。目的:探讨利用CD271磁珠从体外诱导分化的人胚胎干细胞中分选出成骨细胞的可行性。方法:人胚胎干细胞株SYSU-2形成拟胚体6d,贴壁分化14d后,通过CD271磁珠分选技术获得CD271阳性细胞,进一步检测CD271阳性细胞核型,表面抗原表达及分化潜能。结果与结论:CD271阳性细胞与骨髓来源的间充质干细胞形态相似,为长梭形,并可在体外保持稳定核型至14代左右。同时,这种细胞也具有与骨髓来源的间充质干细胞相似的表面抗原标记(CD45阴性和CD73,CD105,CD166阳性)。分化潜能鉴定证明CD271阳性细胞只能诱导形成成骨细胞。这对于研究成骨发育的机制和为骨组织工程提供足量安全有效的种子细胞都有着重要意义。

BACKGROUND： Studies on osteoblasts derived from human embryonic stem cells （hESCs） had made remarkable results, but the induced methods can produce a great number of effective osteoblasts derived from hESCs without other miscellaneous cells is not established. OBJECTIVE： To explore the feasibility of osteoblasts derived from hESCs in vitro by using CD271 magnetic beads sorting system. METHODS： CD271 positive cells were obtained from hESCs lines SYSU-2 cells through embryoid bodies formation at 6 days and attachment differentiation at 14 days by using CD271 magnetic beads sorting technology. Cell surface antigen expression, karyotype analysis and potential of differentiation were detected on CD271 positive cells. RESULTS AND CONCLUSION： CD271 positive cells exhibited regular spindle shape, which was similar to morphology of bone marrow-derived mesenchymal stem cells （BMSCs）, and could be cultured in vitro about 14 generation remaining stable karyotype. At the same time, these cells expressed similar cell surface antigen marks （CD45 negative; CD73, CD105 and CD166 positive） with BMSCs. But differentiation potential identification showed that CD271 positive cells were solely induced osteoblasts formation. It has tremendous significance to study for the bone development mechanism and supply with enough, safe and efficient seed cells for bone tissue engineering.