Laitinen Anita, Lampinen Milla, Liedtke Stefanie, Kilpinen Lotta, Kerkelä Erja, Sarkanen Jertta-Riina, Heinonen Tuula, Kogler Gesine, Laitinen Saara
Cell Therapy Services, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
Faculty of Medicine, Department of Pharmacology, University of Helsinki, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland.
Cytotherapy. 2016 Mar;18(3):423-37. doi: 10.1016/j.jcyt.2015.11.014.
Cord blood (CB) is an attractive source of mesenchymal stromal cells (MSCs) because of its abundant availability and ease of collection. However, the success rate of generating CB-MSCs is low. In this study, our aim was to demonstrate the efficiency of our previously described method to obtain MSCs from CB and further characterize them and to study the effects of different culture conditions on MSCs.
CB-MSC cultures were established in low oxygen (3%) conditions on fibronectin in 10% fetal bovine serum containing culture medium supplemented with combinations of growth factors. Cells were characterized for their adipogenic, osteogenic and chondrogenic differentiation capacity; phenotype; and HOX gene expression profile. The functionality of the cells cultured in different media was tested in vitro with angiogenesis and T-cell proliferation assays.
We demonstrate 87% efficacy in generating MSCs from CB. The established cells had typical MSC characteristics with reduced adipogenic differentiation potential and a unique HOX gene fingerprint. Growth factor-rich medium and a 3% oxygen condition enhanced cell proliferation; however, the growth factor-rich medium had a negative effect on the expression of CD90. Dexamethasone-containing medium improved the capacity of the cells to suppress T-cell proliferation, whereas the cells grown without dexamethasone were more able to support angiogenesis.
Our results demonstrate that the composition of expansion medium is critical for the functionality of MSCs and should always be appropriately defined for each purpose.
脐血（CB）是间充质基质细胞（MSCs）的一个有吸引力的来源，因为其来源丰富且易于采集。然而，生成脐血间充质干细胞的成功率较低。在本研究中，我们的目的是证明我们先前描述的从脐血中获取间充质干细胞的方法的效率，并进一步对其进行表征，以及研究不同培养条件对间充质干细胞的影响。
在低氧（3%）条件下，于含有生长因子组合的10%胎牛血清培养基中，在纤连蛋白上建立脐血间充质干细胞培养体系。对细胞进行脂肪生成、成骨和成软骨分化能力、表型以及HOX基因表达谱的表征。通过血管生成和T细胞增殖试验在体外测试不同培养基中培养的细胞的功能。
我们证明从脐血中生成间充质干细胞的效率为87%。所建立的细胞具有典型的间充质干细胞特征，脂肪生成分化潜能降低且具有独特的HOX基因指纹。富含生长因子的培养基和3%的氧条件可增强细胞增殖；然而，富含生长因子的培养基对CD90的表达有负面影响。含地塞米松的培养基提高了细胞抑制T细胞增殖的能力，而在地塞米松不存在的情况下生长的细胞更能支持血管生成。
我们的结果表明，扩增培养基的组成对间充质干细胞的功能至关重要，并且应始终根据每个目的进行适当定义。
