Hiebl B, Fuhrmann R, Franke R P
Centre for Biomaterial Development, GKSS Research Centre, Teltow, Germany.
Clin Hemorheol Microcirc. 2008;39(1-4):221-8.
Monocytes are broadly discussed in the literature as cells, which can get properties of endothelial progenitor cells after angiogenic stimulation. Angiogenically stimulated monocytes can be used to promote implant vascularisation. A necessity therefore is that these cells can be stored and used after storage without a loose of their characteristic phenotype. In this study we tested, if freshly thawed cryopreserved human monocytes are positive for the mo/macrophage markers CD14 and CD68 and the endothelial marker CD31 after thawing and following angiogenic stimulation in a VEGF-A(165) enriched (10 ng/ml) angiogenic medium. Thereby the monocytes were tested before and after differentiation towards macrophages. The results revealed that freshly thawed human CD14 positive monocytes are positive for CD14, CD68 and CD31 after angiogenic stimulation. This CD specification was much more intense in the differentiated cells. The differentiation step also resulted in an increased cell count. Both results can be attributed to the method of differentiation, were cell culture bags were used instead of common cell culture dishes. Additionally the differentiation medium (X-VIVO 10+10% FCS) was specifically adapted to the requirements of monocytes/macrophages. The study showed that human CD14 positive monocytes can be thawed after cryopreservation without loss of their monocytes/macrophage phenotype and without loss of their ability to get angiogenically stimulated. To enhance the efficiency of both steps (thawing, angiogenic stimulation) it can be useful to differentiate the thawed cells in cell culture bags by the use of X-VIVO 10 (+10% FCS) before angiogenic stimulation.
单核细胞在文献中被广泛讨论为一类细胞，它们在血管生成刺激后可获得内皮祖细胞的特性。经血管生成刺激的单核细胞可用于促进植入物血管化。因此，必要条件是这些细胞能够被储存，并且储存后使用时不会丧失其特征性表型。在本研究中，我们测试了新鲜解冻的低温保存的人单核细胞在解冻后以及在富含VEGF-A(165)（10 ng/ml）的血管生成培养基中进行血管生成刺激后，是否对单核细胞/巨噬细胞标志物CD14和CD68以及内皮标志物CD31呈阳性。由此，在单核细胞向巨噬细胞分化之前和之后对其进行了测试。结果显示，新鲜解冻的人CD14阳性单核细胞在血管生成刺激后对CD14、CD68和CD31呈阳性。这种CD特异性在分化细胞中更为强烈。分化步骤还导致细胞数量增加。这两个结果都可归因于分化方法，即使用细胞培养袋而非普通细胞培养皿。此外，分化培养基（X-VIVO 10 + 10% FCS）是专门根据单核细胞/巨噬细胞的需求进行调整的。该研究表明，人CD14阳性单核细胞在低温保存后解冻时不会丧失其单核细胞/巨噬细胞表型，也不会丧失其接受血管生成刺激的能力。为了提高两个步骤（解冻、血管生成刺激）的效率，在血管生成刺激之前，使用X-VIVO 10（+10% FCS）在细胞培养袋中分化解冻后的细胞可能会有所帮助。
