Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions

Abstract

Pluripotent stem cells (PSCs) are characterized by the ability to self-renew as well as undergo multidirectional differentiation. Culture conditions have a pivotal influence on differentiation pattern. In the current study, we compared the fate of mouse PSCs using two culture media: (1) chemically defined, free of animal reagents, and (2) standard one relying on the serum supplementation. Moreover, we assessed the influence of selected regulators (WNTs, SHH) on PSC differentiation. We showed that the differentiation pattern of PSCs cultured in both systems differed significantly: cells cultured in chemically defined medium preferentially underwent ectodermal conversion while their endo- and mesodermal differentiation was limited, contrary to cells cultured in serum-supplemented medium. More efficient ectodermal differentiation of PSCs cultured in chemically defined medium correlated with higher activity of SHH pathway while endodermal and mesodermal conversion of cells cultured in serum-supplemented medium with higher activity of WNT/JNK pathway. However, inhibition of either canonical or noncanonical WNT pathway resulted in the limitation of endo- and mesodermal conversion of PSCs. In addition, blocking WNT secretion led to the inhibition of PSC mesodermal differentiation, confirming the pivotal role of WNT signaling in this process. In contrast, SHH turned out to be an inducer of PSC ectodermal, not mesodermal differentiation.

Stiffness‐Tunable Hydrogel‐Sandwich Culture Modulates the YAP‐Mediated Mechanoresponse in Induced‐Pluripotent Stem Cell Embryoid Bodies and Augments Cardiomyocyte Differentiation

“…Mouse ESCs spontaneously differentiate into endodermal and mesodermal lineages, while ectodermal differentiation is limited in the presence of serum. [ 39 ] To investigate the effect of HGSCs on mesodermal differentiation, the miPSC‐EBs collected on Day 5 were labeled with an anti‐Brachyury antibody for whole‐mount immunofluorescence staining (Figure 3c). Brachyury‐positive cells were detected on the surface of miPSC‐Ebs.…”

The asymmetric expression of HSPA2 in blastomeres governs the first embryonic cell-fate decision

“…The most significant GO enriched terms were closely related to cell fate commitment, stem cell population maintenance, stem cell differentiation and blastocyst development (Figure 2E). According to KEGG analysis, the downregulated genes were primarily associated with Hippo and Wnt signaling pathways (Figure 2F), which are known to play an important role in cell proliferation, differentiation, and fate decisions in embryonic development [22][23][24][25]. Furthermore, the blastocyst formation rate in the Hspa2-KD group (52.96±2.489%) was significantly lower than that in the NC group (81.63±2.000%) (Figure 2G, H), thus suggesting that HSPA2 plays an essential role in preimplantation embryonic development.…”

A simple tool for the synchronous differentiation of human induced pluripotent stem cells into pancreatic progenitors

“…[14,15] Furthermore, significant variations in the efficiency of differentiation have been observed among different PSC lines, with some lineages differentiating more easily into a particular cell type than others. [16,17] Although the differentiation methods for PSCs have already been well-established, these methods are limited by relatively low differentiation efficiencies and poor reproducibility. [14,18] Therefore, to improve the reliability of this technology, it is essential to develop a robust, highly efficient, homogeneous, and reproducible pancreatic progenitor differentiation system.…”