Ningxia Human Stem Cell Institute, The Affiliated Hospital of Ningxia Medical University, Yinchuan, Ningxia, PR China.
Mol Med Rep. 2012 Oct;6(4):760-6. doi: 10.3892/mmr.2012.1000. Epub 2012 Jul 23.
Human placenta-derived mesenchymal stem cells (P-MSCs) have drawn increasing attention in the field of stem cell research due to their potential in clinical applications as well as their rich and easy to procure cell source. While studies demonstrating the potential of P-MSCs for therapeutic transplantations have been documented, a clinically compliant procedure for P-MSC expansion in vitro has yet to be established. To this end, previous studies have demonstrated that MSCs of bone marrow and cord blood origins cultured in human cord blood serum (hCBS) are comparable to those cultured in fetal bovine serum (FBS), indicating that hCBS may be an alternative to FBS for the development of in vitro cell expansion procedures free of animal components. However, stem cells from origins other than bone marrow or cord blood, particularly from human placental tissues, which have demonstrated a good potential for clinical applications, have not been characterized under similar conditions. In this study, in an attempt to define a clinically compliant protocol for P-MSC expansion in vitro, we examined the effects of human hCBS as a replacement for FBS on cell proliferation capacity, differentiation potential, MSC-specific phenotypic expression and the genetic stability of P-MSCs in cultures. P-MSCs expanded in vitro in autologous hCBS maintained the capacity of self‑renewal and expressed surface antigens characteristic of bone marrow-derived mesenchymal stem cells. Under differentiation conditions, the P-MSCs expanded in hCBS developed into adipogenic, osteogenic and neurogenic cell phenotypes. Chromosomal karyotyping and single cell gel electrophoresis analysis demonstrated that P-MSCs cultured in autologous hCBS were genetically stable. These results suggest that autologous hCBS may be used as an alternative to FBS for the in vitro expansion of P-MSCs for clinical applications.
人胎盘间充质干细胞(P-MSCs)因其在临床应用中的潜力以及丰富且易于获取的细胞来源,在干细胞研究领域引起了越来越多的关注。虽然已经有研究证明了 P-MSCs 用于治疗性移植的潜力,但尚未建立体外 P-MSC 扩增的临床合规程序。为此,先前的研究表明,在人脐血血清(hCBS)中培养的骨髓和脐血来源的 MSC 与在胎牛血清(FBS)中培养的 MSC 相当,这表明 hCBS 可能是替代 FBS 用于开发无动物成分的体外细胞扩增程序的一种选择。然而,除骨髓或脐血以外的来源(特别是具有良好临床应用潜力的人胎盘组织来源)的干细胞尚未在类似条件下进行表征。在这项研究中,为了尝试定义一种临床合规的体外 P-MSC 扩增方案,我们研究了用人 hCBS 替代 FBS 对细胞增殖能力、分化潜能、MSC 特异性表型表达和 P-MSCs 培养中的遗传稳定性的影响。在自体 hCBS 中体外扩增的 P-MSCs 保持了自我更新的能力,并表达了骨髓来源的间充质干细胞的表面抗原。在分化条件下,hCBS 中扩增的 P-MSCs 可分化为成脂肪细胞、成骨细胞和神经细胞表型。染色体核型分析和单细胞凝胶电泳分析表明,在自体 hCBS 中培养的 P-MSCs 遗传稳定。这些结果表明,自体 hCBS 可替代 FBS 用于临床应用的 P-MSCs 体外扩增。