Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells

GÜZEL, SAIME; Yalcin, Abdullah; Gürpınar, Yunus; guler, sabire

doi:10.55730/1300-0144.5725

Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells

Saime GÜZEL, (Bursa Uludağ Üniversitesi, Veteriner Fakültesi, Biyokimya Anabilim Dalı, Bursa, Türkiye)

Abdullah YALÇIN, (Bursa Uludağ Üniversitesi, Veteriner Fakültesi, Biyokimya Anabilim Dalı, Bursa, Türkiye)

Yunus GÜRPINAR, (Koç Üniversitesi, Translasyonel Tıp Araştırma Merkezi, İstanbul, Türkiye)

Sabire GÜLER (Bursa Uludağ Üniversitesi, Veteriner Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Bursa, Türkiye)

Turkish Journal of Medical Sciences

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Yıl: 2023 Cilt: 53 Sayı: 6 Sayfa Aralığı: 1565 - 1573 Metin Dili: İngilizce DOI: 10.55730/1300-0144.5725 İndeks Tarihi: 17-01-2024

Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells

Öz:

Background/aim: Type 1 diabetes mellitus (T1DM) is caused by the autoimmune-mediated destruction of insulin-producing cells (IPCs) and still has no effective cure. Better understanding of the molecular mechanisms involved in the differentiation of embryonic stem cells (ESCs) into IPCs may help us improve the therapeutic strategies for treating T1DM. 6-phosphofructo-2-kinase/fructose-2,6- bisphosphatases (Pfkfb1–4) are key regulators of glucose metabolism. Although Pfkfb3 has been shown to be required for the growth of early differentiated mouse ESCs (mESCs), more studies are needed to further assess the roles of Pfkfb isoenzymes in embryonic development and differentiation, particularly into specific cell types. In this study, we aimed to elucidate the changes in the expression of Pfkfb isoenzymes on the differentiation of mESCs into IPCs. Materials and methods: A 3-step protocol was used to differentiate R1 and J1 mESCs into IPCs. The changes in the gene expression of MafA, MafB, Ins2, and Nkx6.1 (IPC specific markers) and Pfkfb1–4 were analyzed using real-time quantitative polymerase chain reaction (qPCR). Insulin expression and secretion were determined by immunofluorescence (IF) staining and the enzyme linked immunosorbent assay (ELISA), respectively. Results: Upon differentiation, the IPC specific markers in differentiated cells were upregulated. Continued differentiation was confirmed by the development of insulin-positive islet-like clusters that secreted insulin in response to glucose uptake. Expressions of the Pfkfb2 and Pfkfb3 isoenzymes were markedly increased in various stages of differentiation. Conclusion: These findings suggest that Pfkfb2 and Pfkfb3 may impact the differentiation of mESCs into IPCs and the regulation of the insulin response to glucose levels. This study also lays a foundation for researchers to further probe the roles of Pfkfb isoenzymes on the differentiation of mESCs into IPCs and may open new avenues for regenerative medicine.

Anahtar Kelime: Mouse embryonic stem cells insulin-producing cells differentiation 6-phosphofructo-2-kinase/fructose-2 6-bisphosphatase

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	GÜZEL S, Yalcin A, Gürpınar Y, guler s (2023). Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. , 1565 - 1573. 10.55730/1300-0144.5725
Chicago	GÜZEL SAIME,Yalcin Abdullah,Gürpınar Yunus,guler sabire Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. (2023): 1565 - 1573. 10.55730/1300-0144.5725
MLA	GÜZEL SAIME,Yalcin Abdullah,Gürpınar Yunus,guler sabire Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. , 2023, ss.1565 - 1573. 10.55730/1300-0144.5725
AMA	GÜZEL S,Yalcin A,Gürpınar Y,guler s Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. . 2023; 1565 - 1573. 10.55730/1300-0144.5725
Vancouver	GÜZEL S,Yalcin A,Gürpınar Y,guler s Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. . 2023; 1565 - 1573. 10.55730/1300-0144.5725
IEEE	GÜZEL S,Yalcin A,Gürpınar Y,guler s "Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells." , ss.1565 - 1573, 2023. 10.55730/1300-0144.5725
ISNAD	GÜZEL, SAIME vd. "Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells". (2023), 1565-1573. https://doi.org/10.55730/1300-0144.5725

APA	GÜZEL S, Yalcin A, Gürpınar Y, guler s (2023). Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. Turkish Journal of Medical Sciences, 53(6), 1565 - 1573. 10.55730/1300-0144.5725
Chicago	GÜZEL SAIME,Yalcin Abdullah,Gürpınar Yunus,guler sabire Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. Turkish Journal of Medical Sciences 53, no.6 (2023): 1565 - 1573. 10.55730/1300-0144.5725
MLA	GÜZEL SAIME,Yalcin Abdullah,Gürpınar Yunus,guler sabire Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. Turkish Journal of Medical Sciences, vol.53, no.6, 2023, ss.1565 - 1573. 10.55730/1300-0144.5725
AMA	GÜZEL S,Yalcin A,Gürpınar Y,guler s Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. Turkish Journal of Medical Sciences. 2023; 53(6): 1565 - 1573. 10.55730/1300-0144.5725
Vancouver	GÜZEL S,Yalcin A,Gürpınar Y,guler s Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells. Turkish Journal of Medical Sciences. 2023; 53(6): 1565 - 1573. 10.55730/1300-0144.5725
IEEE	GÜZEL S,Yalcin A,Gürpınar Y,guler s "Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells." Turkish Journal of Medical Sciences, 53, ss.1565 - 1573, 2023. 10.55730/1300-0144.5725
ISNAD	GÜZEL, SAIME vd. "Expression of Pfkfb isoenzymes during in vitro differentiation of mouse embryonic stem cells into insulin-producing cells". Turkish Journal of Medical Sciences 53/6 (2023), 1565-1573. https://doi.org/10.55730/1300-0144.5725