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Bile acids induce hepatic differentiation of mesenchymal stem cells.

Sawitza I, Kordes C, Götze S, Herebian D, Häussinger D - Sci Rep (2015)

Bottom Line: Mesenchymal stem cells (MSC) have the potential to differentiate into multiple cell lineages and their therapeutic potential has become obvious.In the liver, MSC are represented by stellate cells which have the potential to differentiate into hepatocytes after stimulation with growth factors.Notch, hedgehog, transforming growth factor-β/bone morphogenic protein family and non-canonical Wnt signalling were also essential for bile acid-mediated differentiation, whereas β-catenin-dependent Wnt signalling was able to attenuate this process.

View Article: PubMed Central - PubMed

Affiliation: Clinic of Gastroenterology, Hepatology and Infectious Diseases.

ABSTRACT
Mesenchymal stem cells (MSC) have the potential to differentiate into multiple cell lineages and their therapeutic potential has become obvious. In the liver, MSC are represented by stellate cells which have the potential to differentiate into hepatocytes after stimulation with growth factors. Since bile acids can promote liver regeneration, their influence on liver-resident and bone marrow-derived MSC was investigated. Physiological concentrations of bile acids such as tauroursodeoxycholic acid were able to initiate hepatic differentiation of MSC via the farnesoid X receptor and transmembrane G-protein-coupled bile acid receptor 5 as investigated with knockout mice. Notch, hedgehog, transforming growth factor-β/bone morphogenic protein family and non-canonical Wnt signalling were also essential for bile acid-mediated differentiation, whereas β-catenin-dependent Wnt signalling was able to attenuate this process. Our findings reveal bile acid-mediated signalling as an alternative way to induce hepatic differentiaion of stem cells and highlight bile acids as important signalling molecules during liver regeneration.

No MeSH data available.


Related in: MedlinePlus

Expression of bile acid receptors and transporters in freshly isolated and TUDCA-treated HSC from rats.Freshly isolated HSC from rats were treated with 2 μM TUDCA for 21 days and the expression of the bile acid receptors (a) Fxr and (b) Tgr5 as well as the bile acid transporters (c) Asbt, (d) Oatp4, (e) Bsep and (f) Ntcp was investigated by qPCR in weekly intervals (n = 3). HSC of the control received the same medium but without TUDCA. The expression of (a) Fxr, (c) Asbt, (d) Oatp4 and (e) Bsep mRNA was already detectable in freshly isolated HSC (day 1) but increased significantly in response to TUDCA treatment. (b) The mRNA of Tgr5 remained undetectable in freshly isolated HSC but appeared during culture of HSC under control conditions and after TUDCA treatment. (f) Ntcp expression was not found in freshly isolated HSC and under control conditions, but was induced in response to TUDCA administration and served as an additional marker for hepatic differentiation.
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f5: Expression of bile acid receptors and transporters in freshly isolated and TUDCA-treated HSC from rats.Freshly isolated HSC from rats were treated with 2 μM TUDCA for 21 days and the expression of the bile acid receptors (a) Fxr and (b) Tgr5 as well as the bile acid transporters (c) Asbt, (d) Oatp4, (e) Bsep and (f) Ntcp was investigated by qPCR in weekly intervals (n = 3). HSC of the control received the same medium but without TUDCA. The expression of (a) Fxr, (c) Asbt, (d) Oatp4 and (e) Bsep mRNA was already detectable in freshly isolated HSC (day 1) but increased significantly in response to TUDCA treatment. (b) The mRNA of Tgr5 remained undetectable in freshly isolated HSC but appeared during culture of HSC under control conditions and after TUDCA treatment. (f) Ntcp expression was not found in freshly isolated HSC and under control conditions, but was induced in response to TUDCA administration and served as an additional marker for hepatic differentiation.

Mentions: The molecular mechanisms underlying the TUDCA-mediated hepatic differentiation of MSC were investigated by the use of isolated bmMSC from Fxr and Tgr5 knockout (KO) mice. The knockout of Fxr and Tgr5, respectively, was verified by qPCR in comparison with bmMSC from the corresponding wild type mouse strains. Fxr and Tgr5 expression was only found in bmMSC from wild type control mice (not shown). The deletion of either Fxr (Fig. 4a–c) or Tgr5 (Fig. 4d–f) significantly inhibited the TUDCA-mediated hepatic differentiation of bmMSC as indicated by decreased albumin expression and bile acid production when compared to the results obtained with bmMSC from wild type mouse strains. This suggests that both bile acid receptors are involved in the hepatic differentiation induced by bile acids. The requirement of Tgr5 for the initiation of this process was also observed in primary cultures of isolated HSC from Tgr5-KO mice. HSC without Tgr5 showed no significant differentiation after TUDCA treatment, whereas HSC from wild type mice developed into hepatocyte-like cells as indicated by their albumin release (supplemental Fig. S4). However, growth factor-mediated hepatic differentiation was found to be independent from bile acid receptors, since the differentiation of MSC into hepatocyte-like cells was unaffected in bmMSC from Fxr and Tgr5 knockout mice during treatment with hepatocyte growth factor (HGF) and fibroblast growth factor 4 (FGF4) (Fig. 4g–l). The expression of Fxr and Tgr5 was found to be up-regulated early during culture of rat HSC as investigated by qPCR (Fig. 5a,b), immunofluorescence and Western blot analysis (Fig. 6a–f). After 4 days in culture, Fxr became clearly detectable in the cell nuclei and Tgr5 appeared in the cell membrane of rat HSC under serum-free conditions (Fig. 6a–f), indicating that only activated HSC can properly respond to bile acids. The expression of Fxr and Tgr5 strongly increased in HSC when TUDCA was added to the culture medium (Fig. 5a,b). Also bile acid transporters such as the apical sodium-dependent bile acid transporter (Asbt), organic anion-transporting polypeptide 4 (Oatp4) and bile salt export pump (Bsep) were stronger expressed in response to TUDCA treatment (Fig. 5c–e, supplemental Fig. S5a–f). Although Asbt and Oatp4 were found to be expressed in freshly isolated HSC as investigated at the mRNA level, only small amounts of these transporters were detectable at protein level by immunofluorescence staining (supplemental Fig. S5a–h), suggesting that quiescent HSC can take up bile acids at least in principle. The expression of Bsep remained weak and Ntcp was undetectable in freshly isolated rat HSC as analysed by qPCR (Fig. 5e,f). The hepatocyte-specific Ntcp remained undetectable in HSC without TUDCA treatment (control), but was induced after application of TUDCA, as another indication for their differentiation into hepatocyte-like cells (Fig. 5f; supplemental Fig. S5g, h).


Bile acids induce hepatic differentiation of mesenchymal stem cells.

Sawitza I, Kordes C, Götze S, Herebian D, Häussinger D - Sci Rep (2015)

Expression of bile acid receptors and transporters in freshly isolated and TUDCA-treated HSC from rats.Freshly isolated HSC from rats were treated with 2 μM TUDCA for 21 days and the expression of the bile acid receptors (a) Fxr and (b) Tgr5 as well as the bile acid transporters (c) Asbt, (d) Oatp4, (e) Bsep and (f) Ntcp was investigated by qPCR in weekly intervals (n = 3). HSC of the control received the same medium but without TUDCA. The expression of (a) Fxr, (c) Asbt, (d) Oatp4 and (e) Bsep mRNA was already detectable in freshly isolated HSC (day 1) but increased significantly in response to TUDCA treatment. (b) The mRNA of Tgr5 remained undetectable in freshly isolated HSC but appeared during culture of HSC under control conditions and after TUDCA treatment. (f) Ntcp expression was not found in freshly isolated HSC and under control conditions, but was induced in response to TUDCA administration and served as an additional marker for hepatic differentiation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4548444&req=5

f5: Expression of bile acid receptors and transporters in freshly isolated and TUDCA-treated HSC from rats.Freshly isolated HSC from rats were treated with 2 μM TUDCA for 21 days and the expression of the bile acid receptors (a) Fxr and (b) Tgr5 as well as the bile acid transporters (c) Asbt, (d) Oatp4, (e) Bsep and (f) Ntcp was investigated by qPCR in weekly intervals (n = 3). HSC of the control received the same medium but without TUDCA. The expression of (a) Fxr, (c) Asbt, (d) Oatp4 and (e) Bsep mRNA was already detectable in freshly isolated HSC (day 1) but increased significantly in response to TUDCA treatment. (b) The mRNA of Tgr5 remained undetectable in freshly isolated HSC but appeared during culture of HSC under control conditions and after TUDCA treatment. (f) Ntcp expression was not found in freshly isolated HSC and under control conditions, but was induced in response to TUDCA administration and served as an additional marker for hepatic differentiation.
Mentions: The molecular mechanisms underlying the TUDCA-mediated hepatic differentiation of MSC were investigated by the use of isolated bmMSC from Fxr and Tgr5 knockout (KO) mice. The knockout of Fxr and Tgr5, respectively, was verified by qPCR in comparison with bmMSC from the corresponding wild type mouse strains. Fxr and Tgr5 expression was only found in bmMSC from wild type control mice (not shown). The deletion of either Fxr (Fig. 4a–c) or Tgr5 (Fig. 4d–f) significantly inhibited the TUDCA-mediated hepatic differentiation of bmMSC as indicated by decreased albumin expression and bile acid production when compared to the results obtained with bmMSC from wild type mouse strains. This suggests that both bile acid receptors are involved in the hepatic differentiation induced by bile acids. The requirement of Tgr5 for the initiation of this process was also observed in primary cultures of isolated HSC from Tgr5-KO mice. HSC without Tgr5 showed no significant differentiation after TUDCA treatment, whereas HSC from wild type mice developed into hepatocyte-like cells as indicated by their albumin release (supplemental Fig. S4). However, growth factor-mediated hepatic differentiation was found to be independent from bile acid receptors, since the differentiation of MSC into hepatocyte-like cells was unaffected in bmMSC from Fxr and Tgr5 knockout mice during treatment with hepatocyte growth factor (HGF) and fibroblast growth factor 4 (FGF4) (Fig. 4g–l). The expression of Fxr and Tgr5 was found to be up-regulated early during culture of rat HSC as investigated by qPCR (Fig. 5a,b), immunofluorescence and Western blot analysis (Fig. 6a–f). After 4 days in culture, Fxr became clearly detectable in the cell nuclei and Tgr5 appeared in the cell membrane of rat HSC under serum-free conditions (Fig. 6a–f), indicating that only activated HSC can properly respond to bile acids. The expression of Fxr and Tgr5 strongly increased in HSC when TUDCA was added to the culture medium (Fig. 5a,b). Also bile acid transporters such as the apical sodium-dependent bile acid transporter (Asbt), organic anion-transporting polypeptide 4 (Oatp4) and bile salt export pump (Bsep) were stronger expressed in response to TUDCA treatment (Fig. 5c–e, supplemental Fig. S5a–f). Although Asbt and Oatp4 were found to be expressed in freshly isolated HSC as investigated at the mRNA level, only small amounts of these transporters were detectable at protein level by immunofluorescence staining (supplemental Fig. S5a–h), suggesting that quiescent HSC can take up bile acids at least in principle. The expression of Bsep remained weak and Ntcp was undetectable in freshly isolated rat HSC as analysed by qPCR (Fig. 5e,f). The hepatocyte-specific Ntcp remained undetectable in HSC without TUDCA treatment (control), but was induced after application of TUDCA, as another indication for their differentiation into hepatocyte-like cells (Fig. 5f; supplemental Fig. S5g, h).

Bottom Line: Mesenchymal stem cells (MSC) have the potential to differentiate into multiple cell lineages and their therapeutic potential has become obvious.In the liver, MSC are represented by stellate cells which have the potential to differentiate into hepatocytes after stimulation with growth factors.Notch, hedgehog, transforming growth factor-β/bone morphogenic protein family and non-canonical Wnt signalling were also essential for bile acid-mediated differentiation, whereas β-catenin-dependent Wnt signalling was able to attenuate this process.

View Article: PubMed Central - PubMed

Affiliation: Clinic of Gastroenterology, Hepatology and Infectious Diseases.

ABSTRACT
Mesenchymal stem cells (MSC) have the potential to differentiate into multiple cell lineages and their therapeutic potential has become obvious. In the liver, MSC are represented by stellate cells which have the potential to differentiate into hepatocytes after stimulation with growth factors. Since bile acids can promote liver regeneration, their influence on liver-resident and bone marrow-derived MSC was investigated. Physiological concentrations of bile acids such as tauroursodeoxycholic acid were able to initiate hepatic differentiation of MSC via the farnesoid X receptor and transmembrane G-protein-coupled bile acid receptor 5 as investigated with knockout mice. Notch, hedgehog, transforming growth factor-β/bone morphogenic protein family and non-canonical Wnt signalling were also essential for bile acid-mediated differentiation, whereas β-catenin-dependent Wnt signalling was able to attenuate this process. Our findings reveal bile acid-mediated signalling as an alternative way to induce hepatic differentiaion of stem cells and highlight bile acids as important signalling molecules during liver regeneration.

No MeSH data available.


Related in: MedlinePlus