To cure the epidemic of diabetes, in vitro produced β-like cells are lauded for cell therapy of diabetic patients. In this regard, we investigated the effects of different concentrations of bone morphogenetic protein 7 (BMP7) on the differentiation of rat pancreatic ductal epithelial-like stem cells (PDESCs) into β-like cells. For inducement of the differentiation, PDESCs were cultured in the basal media (H-DMEM + 10 % FBS + 1% penicillin-streptomycin) supplemented with 5 ng/mL, 10 ng/mL, 15 ng/mL, and 20 ng/mL of BMP7 for 28 days. To corroborate the identity of induced cells, they were examined through cell morphology, dithizone (DTZ) staining, immunofluorescence staining, real-time polymerase chain reaction (qPCR), and glucose-stimulated insulin secretion assay (GSIS). The enrichment of induced cells was high among 5 ng/mL, 10 ng/mL, 15 ng/mL, and 20 ng/mL of BMP7 supplemented groups as compared to the control group. Further, the induced cells were positive, while, the control group cells were negative to DTZ staining and the differentiated cells also have shown the upregulated expression of β cell-specific marker genes (Ins1 and Pdx1). The GSIS assay of inducement groups for insulin and C-peptide secretion has shown significantly higher values as compared to the control group (P < 0.01). Hence, the addition of BMP7 to basal medium has effectually induced differentiation of adult rat PDESCs into islet like-cell clusters containing insulin-secreting β-like cells.

在体外治愈糖尿病的流行产生的 β 样细胞因用于糖尿病患者的细胞治疗而受到称赞。在这方面，我们研究了不同浓度的骨形态发生蛋白 7 (BMP7) 对大鼠胰腺导管上皮样干细胞 (PDESC) 向 β 样细胞分化的影响。为了诱导分化，PDESCs 在基础培养基（H-DMEM + 10% FBS + 1% 青霉素-链霉素）中培养，并辅以 5 ng/mL、10 ng/mL、15 ng/mL 和 20 ng/mL BMP7 28 天。为了证实诱导细胞的身份，通过细胞形态学、双硫腙 (DTZ) 染色、免疫荧光染色、实时聚合酶链反应 (qPCR) 和葡萄糖刺激胰岛素分泌测定 (GSIS) 对它们进行了检查。诱导细胞的富集在 5 ng/mL、10 ng/mL、15 ng/mL、和 20 ng/mL 的 BMP7 补充组与对照组相比。此外，诱导细胞呈阳性，而对照组细胞对 DTZ 染色呈阴性，分化细胞也显示出 β 细胞特异性标记基因（Ins1 和 Pdx1）的表达上调。与对照组相比，胰岛素和 C 肽分泌诱导组的 GSIS 测定显示出显着更高的值 (P < 0.01)。因此，将 BMP7 添加到基础培养基中有效地诱导了成年大鼠 PDESC 分化为含有分泌胰岛素的 β 样细胞的胰岛样细胞簇。对照组细胞对 DTZ 染色呈阴性，分化细胞也显示出 β 细胞特异性标记基因（Ins1 和 Pdx1）的上调表达。与对照组相比，胰岛素和 C 肽分泌诱导组的 GSIS 测定显示出显着更高的值 (P < 0.01)。因此，将 BMP7 添加到基础培养基中有效地诱导了成年大鼠 PDESC 分化为含有分泌胰岛素的 β 样细胞的胰岛样细胞簇。对照组细胞对 DTZ 染色呈阴性，分化细胞也显示出 β 细胞特异性标记基因（Ins1 和 Pdx1）的上调表达。与对照组相比，胰岛素和 C 肽分泌诱导组的 GSIS 测定显示出显着更高的值 (P < 0.01)。因此，将 BMP7 添加到基础培养基中有效地诱导了成年大鼠 PDESC 分化为含有分泌胰岛素的 β 样细胞的胰岛样细胞簇。