Ayala-Grosso Carlos, Pieruzzini Rosalinda, Vargas-Saturno Leslie, Cardier José E
Unidad de Terapia Celular, Laboratorio de Patología Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela.
Servicio de Neurorinología, Departamento de Otorrinolaringología, Hospital Militar Dr. Carlos Arvelo, Caracas, Venezuela.
Biomedica. 2020 Mar 1;40(1):72-88. doi: 10.7705/biomedica.4762.
The olfactory neuro-epithelium has an intrinsic capability of renewal during lifetime provided by the existence of globose and horizontal olfactory precursor cells. Additionally, mesenchymal stromal olfactory cells also support the homeostasis of the olfactory mucosa cell population. Under in vitro culture conditions with Dulbecco modified eagle/F12 medium supplemented with 10% fetal bovine serum, tissue biopsies from upper turbinate have generated an adherent population of cells expressing mainly mesenchymal stromal phenotypic markers. A closer examination of these cells has also found co-expression of olfactory precursors and ensheathing cell phenotypic markers. These results were suggestive of a unique property of olfactory mesenchymal stromal cells as potentially olfactory progenitor cells.
To study whether the expression of these proteins in mesenchymal stromal cells is modulated upon neuronal differentiation.
We observed the phenotype of olfactory stromal cells under DMEM/F12 plus 10% fetal bovine serum in comparison to cells from spheres induced by serum-free medium plus growth factors inducers of neural progenitors.
The expression of mesenchymal stromal (CD29+, CD73+, CD90+, CD45-), horizontal basal (ICAM-1/CD54+, p63+, p75NGFr+), and ensheathing progenitor cell (nestin+, GFAP+) proteins was determined in the cultured population by flow cytometry. The determination of Oct 3/4, Sox-2, and Mash-1 transcription factors, as well as the neurotrophins BDNF, NT3, and NT4 by RT-PCR in cells, was indicative of functional heterogeneity of the olfactory mucosa tissue sample.
Mesenchymal and olfactory precursor proteins were downregulated by serum-free medium and promoted differentiation of mesenchymal stromal cells into neurons and astroglial cells.
嗅神经上皮在其生命周期内具有自我更新的内在能力,这得益于球状和水平嗅前体细胞的存在。此外,间充质基质嗅细胞也支持嗅黏膜细胞群体的稳态。在补充有10%胎牛血清的杜尔贝科改良伊格尔/F12培养基的体外培养条件下,取自上鼻甲的组织活检样本产生了一群主要表达间充质基质表型标志物的贴壁细胞。对这些细胞的进一步检查还发现了嗅前体和包被细胞表型标志物的共表达。这些结果提示嗅间充质基质细胞具有作为潜在嗅祖细胞的独特特性。
研究这些蛋白质在间充质基质细胞中的表达在神经元分化时是否受到调节。
我们将在DMEM/F12加10%胎牛血清条件下培养的嗅基质细胞的表型与由无血清培养基加神经祖细胞生长因子诱导剂形成的球体中的细胞表型进行了比较。
通过流式细胞术测定了培养群体中间充质基质(CD29+、CD73+、CD90+、CD45-)、水平基底(ICAM-1/CD54+、p63+、p75NGFr+)和包被祖细胞(巢蛋白+、GFAP+)蛋白的表达。通过RT-PCR测定细胞中Oct 3/4、Sox-2和Mash-1转录因子以及神经营养因子BDNF、NT3和NT4的表达,表明嗅黏膜组织样本具有功能异质性。
无血清培养基下调了间充质和嗅前体蛋白,并促进间充质基质细胞向神经元和星形胶质细胞分化。