Berlin-Brandenburg Center for Regenerative Therapies, Charité-University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
J Transl Med. 2013 Sep 27;11:236. doi: 10.1186/1479-5876-11-236.
Clinical cardiac cell therapy using autologous somatic stem cells is restricted by age and disease-associated impairment of stem cell function. Juvenile cells possibly represent a more potent alternative, but the impact of patient-related variables on such cell products is unknown. We therefore evaluated the behavior of neonatal cord blood mesenchymal stem cells (CB-MSC) in the presence of serum from patients with advanced heart failure (HF).
Human serum was obtained from patients with severe HF (n = 21) and from healthy volunteers (n = 12). To confirm the systemic quality of HF in the sera, TNF-α and IL-6 were quantified. CB-MSC from healthy neonates were cultivated for up to 14 days in medium supplemented with 10% protein-normalized human HF or control serum or fetal calf serum (FCS).
All HF sera contained increased cytokine concentrations (IL-6, TNF-α). When exposed to HF serum, CB-MSC maintained basic MSC properties as confirmed by immunophenotyping and differentiation assays, but clonogenic cells were reduced in number and gave rise to substantially smaller colonies in the CFU-F assay. Cell cycle analysis pointed towards G1 arrest. CB-MSC metabolic activity and proliferation were significantly impaired for up to 3 days as measured by MTS turnover, BrdU incorporation and DAPI + nuclei counting. On day 5, however, CB-MSC growth kinetics approached control serum levels, though protein expression of cell cycle inhibitors (p21, p27), and apoptosis marker Caspase 3 remained elevated. Signal transduction included the stress and cytokine-induced JNK and ERK1/2 MAP kinase pathways.
Heart failure temporarily inhibits clonality and proliferation of "healthy" juvenile MSC in vitro. Further studies should address the in vivo and clinical relevance of this finding.
使用自体体细胞的临床心脏细胞疗法受到年龄和与疾病相关的干细胞功能障碍的限制。幼年细胞可能代表一种更有效的替代方法，但患者相关变量对这种细胞产品的影响尚不清楚。因此，我们评估了在存在晚期心力衰竭（HF）患者血清的情况下，新生脐带血间充质干细胞（CB-MSC）的行为。
从严重 HF 患者（n=21）和健康志愿者（n=12）中获得人血清。为了确认血清中 HF 的系统性质量，定量了 TNF-α和 IL-6。将来自健康新生儿的 CB-MSC 在补充有 10％蛋白归一化的 HF 或对照血清或胎牛血清（FCS）的培养基中培养长达 14 天。
所有 HF 血清均含有较高的细胞因子浓度（IL-6，TNF-α）。当暴露于 HF 血清时，CB-MSC 保持基本的 MSC 特性，这通过免疫表型和分化测定得到证实，但集落形成细胞的数量减少，并且在 CFU-F 测定中形成的集落要小得多。细胞周期分析表明 G1 期停滞。通过 MTS 转换，BrdU 掺入和 DAPI +细胞核计数测量，CB-MSC 的代谢活性和增殖在长达 3 天的时间内受到显著抑制。然而，在第 5 天，CB-MSC 的生长动力学接近对照血清水平，尽管细胞周期抑制剂（p21、p27）和凋亡标志物 Caspase 3 的蛋白表达仍然升高。信号转导包括应激和细胞因子诱导的 JNK 和 ERK1/2 MAP 激酶途径。
心力衰竭暂时抑制了“健康”幼年 MSC 的克隆性和增殖。进一步的研究应该解决这种发现的体内和临床相关性。
