Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Biotech Histochem. 2020 Apr;95(3):176-185. doi: 10.1080/10520295.2019.1656345. Epub 2019 Oct 7.
Adipose derived stem cells (ADSCs) are multipotent and can transdifferentiate into neural stem cells. We investigated the transdifferentiation of ADSCs to neural phenotype (NP) cells using selegiline and two-dimensional electrophoresis (2-DE). The perinephric and inguinal fat of rats was collected and used to isolate ADSCs that were characterized by immunophenotyping using flow cytometry. The ADSCs were differentiated into osteogenic and lipogenic cells. The NP cells were generated using 10 mM selegiline and characterized by immunocytochemical staining of nestin and neurofilament 68 (NF-68), and by qRT-PCR of nestin, neurod1 and NF68. Total protein of ADSCs and NP cells was extracted and their proteome patterns were examined using 2-DE. ADSCs carried CD73, CD44 and CD90 cell markers, but not CD34. ADSCs were differentiated into osteocyte and adipocyte lineages. The differentiated NP cells expressed nestin, neuro d1 and NF-68. The proteome pattern of ADSCs was compared with that of NP cells and eight spots showed more than a two fold increase in protein expression. The molecular weights and isoelectric points of these highly expressed proteins were estimated using Melanie software. We compared these results with those of the mouse proteomic database using the protein isoelectric point database, and the functions of the eight proteins in differentiation of NP cells were predicted using the UniProt database. The probable identities of the proteins that showed higher expression in NP cells included cholinesterase, GFRa2, protein kinase C (PKC-eta) and RING finger protein 121. The sequences of the proteins identified from mouse database were aligned by comparing them with similar proteins in rat database using the Basic Local Alignment Search Tool (BLAST). The E values of all aligned proteins were zero, which indicates consistency of the matched protein. These proteins participate in differentiation of the neuron and their overexpression causes ADSCs transdifferentiation into NP cells.
脂肪来源的干细胞（ADSCs）是多能的，可以转分化为神经干细胞。我们使用司来吉兰和二维电泳（2-DE）研究 ADSC 向神经表型（NP）细胞的转分化。从大鼠肾周和腹股沟脂肪中收集并用于分离 ADSC，通过流式细胞术对其进行免疫表型鉴定。ADSC 被分化为成骨细胞和脂肪细胞。使用 10 mM 司来吉兰生成 NP 细胞，并通过巢蛋白和神经丝 68（NF-68）的免疫细胞化学染色以及巢蛋白、神经源性分化 1（neurod1）和 NF68 的 qRT-PCR 进行鉴定。提取 ADSC 和 NP 细胞的总蛋白，并使用 2-DE 检查其蛋白质组图谱。ADSC 携带 CD73、CD44 和 CD90 细胞标志物，但不携带 CD34。ADSC 分化为成骨细胞和脂肪细胞谱系。分化的 NP 细胞表达巢蛋白、神经源性分化 1 和 NF-68。ADSC 的蛋白质组图谱与 NP 细胞的蛋白质组图谱进行比较，有 8 个斑点的蛋白质表达增加了两倍以上。使用 Melanie 软件估计这些高表达蛋白的分子量和等电点。我们使用蛋白质等电点数据库将这些结果与小鼠蛋白质组数据库进行比较，并使用 UniProt 数据库预测这 8 种蛋白质在 NP 细胞分化中的功能。在 NP 细胞中表达较高的蛋白质的可能身份包括胆碱酯酶、GFRa2、蛋白激酶 C（PKC-eta）和环指蛋白 121。使用基本局部比对搜索工具（BLAST）将从小鼠数据库中鉴定的蛋白质的序列与大鼠数据库中的相似蛋白质进行比较，从而对齐它们的序列。所有对齐蛋白的 E 值均为零，这表明匹配蛋白具有一致性。这些蛋白质参与神经元的分化，它们的过度表达导致 ADSC 向 NP 细胞转分化。
