Centre for Stem Cell and Gene Therapies Research and Practice, Kocaeli University, Kocaeli 41380, Turkey.
J Tissue Eng Regen Med. 2011 Jun;5(6):491-500. doi: 10.1002/term.342. Epub 2010 Dec 9.
The direct co-culturing effect of rat bone-marrow-derived mesenchymal stem cells (rBM-MSCs) on the pancreatic-islets (PIs) was studied to obtain functional islet cells. MSCs were isolated from rat bone marrow and cultivated under standard conditions. Following their characterization, the rBM-MSCs were directly (with cell-islet contact) co-cultured with recovered PIs together with the single cell cultures of those cell cultures as a control. The effect of direct co-cultures of rBM-MSCs with the PIs of normal rats was investigated using immunophenotypical and functional methods. The change in the amount of insulin secretion was evaluated as an indicator for differentiation of rBM-MSCs. One approache for in vitro differentiation to achieve reprogramming for differentiation into suitable cell types by changing the microenvironment of the cells to provide signals that might activate metabolic pathways is to use co-cultures with the microenvironment of the specific cells of the desired cell type, tissue/organ extracts, extracellular matrix compounds or biologically absorbable materials. Differentiated rBM-MSCs were found to be immunopositive for the specific insulin-producing cell marker, insulin, but not in undifferentiated rBM-MSCs. The functionality tests by ELISA confirmed that insulin secretion of co-cultured MSCs with islets was higher than that of islets. These evidences indicated that PIs could be regarded as critical components of the stem cell niche, such that MSCs can be differentiated into insulin-producing cells (IPCs). Moreover, direct cell-to-cell contact might provide additional and independent support. This approach would circumvent the need for PI-stem cell co-culture and could potentially facilitate the production of functional IPCs for future clinical applications.
为了获得具有功能的胰岛细胞，研究了大鼠骨髓间充质干细胞（rBM-MSCs）对胰岛（PIs）的直接共培养作用。从大鼠骨髓中分离出 MSC，并在标准条件下进行培养。在对其进行特征描述后，将 rBM-MSCs 与回收的 PIs 直接（细胞-胰岛接触）共培养，同时作为对照，对这些细胞培养物的单细胞培养物进行共培养。使用免疫表型和功能方法研究了 rBM-MSCs 与正常大鼠 PIs 的直接共培养的作用。将胰岛素分泌量的变化评估为 rBM-MSCs 分化的指标。一种体外分化的方法是通过改变细胞的微环境来提供可能激活代谢途径的信号，从而实现重编程为合适的细胞类型，即将细胞与所需细胞类型的特定细胞的微环境、组织/器官提取物、细胞外基质化合物或生物可吸收材料进行共培养。研究发现，分化的 rBM-MSCs 对特定的胰岛素产生细胞标志物胰岛素呈免疫阳性，但未分化的 rBM-MSCs 则不然。通过 ELISA 进行的功能测试证实，与胰岛共培养的 MSC 的胰岛素分泌量高于胰岛。这些证据表明，PIs 可以被视为干细胞龛的关键组成部分，使得 MSC 可以分化为产生胰岛素的细胞（IPCs）。此外，细胞间直接接触可能提供额外且独立的支持。这种方法将避免对 PI-干细胞共培养的需求，并可能为未来的临床应用促进功能性 IPC 的产生提供便利。
