Both BMP4 and serum have significant roles in differentiation of embryonic stem cells to primitive and definitive endoderm

Abstract

Differentiation of embryonic stem (ES) cells is a heterogeneous process which is influenced by different parameters, including growth and differentiation factors. The aim of the present study was to investigate the effect of bone morphogenetic protein-4 (BMP4) signaling on differentiation of mouse ES cells to endodermal lineages. For this purpose, differentiation of the ES cells was induced by embryoid body (EB) formation through hanging drop method. During the suspension stage, EBs were treated with BMP4 in a medium containing either fetal bovine serum (FBS) or knockout serum replacement (KoSR). After plating, EBs showed differentiation to a heterogeneous population of specialized cell types. Two weeks after plating, all the experimental groups expressed three germ layer markers and some primitive and definitive endoderm-specific genes. Quantitative real-time PCR analysis showed higher expression levels of Sox17, Pdx1, Cdx2 and Villin mRNAs in the KoSR plus BMP4 condition and higher Gata4 and Afp expression levels in the FBS plus BMP4 condition. Formation of visceral endoderm and derivatives of definitive endoderm was detected in the BMP4 treated EBs. In conclusion, we demonstrated that both BMP4 signaling and serum composition have significant roles in differentiation of mouse ES cells towards endodermal lineages.

The secretome of MUSE cells contains factors that may play a role in regulation of stemness, apoptosis and immunomodulation

“…They are involved in heightening stem differentiation potential and control cell commitment. [54][55][56] A huge number of transcription factors were identified with IPA and Venn analysis as potential common upstream regulators of protein products evidenced in MSC and Muse cell secretomes. Their activities cover all the aspect of cell biology and it is difficult to relate a single factor with a specific biological activity that could be ascribed to proteins present in secretomes.…”

Two-photon polymerized “nichoid” substrates maintain function of pluripotent stem cells when expanded under feeder-free conditions

“…As expected, the differentiation was negative at day 3 (i.e., with LIF conditioning), but surprisingly it was upregulated at day 21 (i.e., without LIF conditioning). This could be due to the FBS in culture that might contain inductive factors [ 27 ].…”

On the right brain cell track

“…Since then several neural differentiation protocols were developed (e.g. [111][112][113][114][115]), which are mainly based on serum deprivation, addition of ATRA, addition of growth factors or a combination thereof [116]. The model used in this thesis is the mouse embryonic neural stem cell test (mESTn), which was based on protocols developed by Okabe et al and Bibel et al [109,112,117].…”

“…For example, neural crest cells are both present in mESTn and mESTc (chapter 4; [111]), and mESTn and hNPT both contain NPC, neurons and astrocytes (chapter 5, 7, 8). In the future, more in vitro tests with complementary biological domains can be linked up with each other to create a testing battery, for example an endodermal differentiation test alongside mESTc and mESTn to cover early cell differentiation along the three germlines [112,113] and regionally defined neural cultures with multiple cell types for later neural differentiation [23,85,114,115].…”