Tara S, Krishnan Lissy K
Division of Thrombosis Research, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences & Technology, Trivandrum, 695012, India.
J Cell Commun Signal. 2019 Mar;13(1):27-38. doi: 10.1007/s12079-018-0467-1. Epub 2018 May 31.
Isolation of progenitors with regenerative potential and their in vitro induction to specific lineage may be necessary for effective cell transplantation outcome. Earlier, we standardized specific niche for derivation of neural progenitor cells (NPCs) from circulating mononuclear cells to neural like cells (NLC) in vitro, for applications in neural regeneration. The current study analysed the prospective involvement of signaling mechanism for in vitro lineage commitment of circulating NPCs. Preferred mechanism selected was Wnt-like signaling because this is one of the pathways implicated in the central nervous system (CNS) development and homeostasis. We sought to determine the activation of Wnt3a-specific genes in the standardized NPC culture system. To start with, it was found that when standardized NPC culture niche was supplemented with Wnt 3a protein, no additional morphological changes happen. Chemical inhibitors of the pathway retarded NPC to NLC conversion both in the absence and presence of supplemented Wnt-3a. In earlier studies, involvement of the niche constituents- fibronectin (FN), laminin (La) and fibrin (Fib)- for NPC growth and differentiation was established. In an attempt to study the role of these adhesive proteins by adding antibodies against FN, La & Fib together, molecular level signaling changes seen were comparable to that occur in response to Wnt3a chemical inhibitor. Therefore, induction of Wnt 3a-like signal from the matrix-dependent niche constituents may be implicated in the differentiation of NPC to NLC. The results substantiate the potential applications of the fibrin-based composite niche in neural engineering for regeneration.
分离具有再生潜力的祖细胞并在体外将其诱导分化为特定谱系，对于有效的细胞移植结果可能是必要的。此前，我们已标准化了一种特定的微环境，用于在体外将循环单核细胞衍生为神经祖细胞（NPCs），再分化为神经样细胞（NLCs），以应用于神经再生。当前研究分析了循环NPCs体外谱系定向中信号传导机制的潜在作用。选择的首选机制是Wnt样信号传导，因为这是参与中枢神经系统（CNS）发育和稳态的途径之一。我们试图确定标准化NPC培养系统中Wnt3a特异性基因的激活情况。首先发现，当在标准化NPC培养微环境中添加Wnt 3a蛋白时，未发生额外的形态变化。该信号通路的化学抑制剂在添加和未添加Wnt-3a的情况下均阻碍了NPC向NLC的转化。在早期研究中，已证实微环境成分——纤连蛋白（FN）、层粘连蛋白（La）和纤维蛋白（Fib）——对NPC生长和分化的作用。为了通过共同添加针对FN、La和Fib的抗体来研究这些黏附蛋白的作用，观察到的分子水平信号变化与Wnt3a化学抑制剂所引起的变化相当。因此，来自依赖基质的微环境成分的Wnt 3a样信号诱导可能与NPC向NLC的分化有关。这些结果证实了基于纤维蛋白的复合微环境在神经工程再生中的潜在应用。
