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Proteomic profiles of mesenchymal stem cells induced by a liver differentiation protocol.

Leelawat K, Narong S, Chaijan S, Sa-Ngiamsuntorn K, Disthabanchong S, Wongkajornsilp A, Hongeng S - Int J Mol Sci (2010)

Bottom Line: We demonstrated that MSCs treated with the liver differentiation protocol expressed significantly more albumin, CK19 and CK20, than did undifferentiated cells.In addition the results of proteomic study demonstrated increases expression of FEM1B, PSMC2 and disulfide-isomerase A3 in MSCs treated with the liver differentiation protocol.These results from proteomic profiling will not only provide insight into the global responses of MSCs to hepatocyte differentiation, but will also lead to in-depth studies on the mechanisms of proteomic changes in MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Rajavithi Hospital, Rajathevi, Bangkok, 10400, Thailand; E-Mails: kawin.leelawat@gmail.com (K.L.); sirilucknarong@hotmail.com (S.N.); bubu_b600@hotmail.com (S.C.).

ABSTRACT
The replacement of disease hepatocytes and the stimulation of endogenous or exogenous regeneration by human mesenchymal stem cells (MSCs) are promising candidates for liver-directed cell therapy. In this study, we isolated MSCs from adult bone marrow by plastic adhesion and induced differentiation with a liver differentiation protocol. Western blot analyses were used to assess the expression of liver-specific markers. Next, MSC-specific proteins were analyzed with two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). To confirm the results from the proteomic study, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analyses were performed. We demonstrated that MSCs treated with the liver differentiation protocol expressed significantly more albumin, CK19 and CK20, than did undifferentiated cells. In addition the results of proteomic study demonstrated increases expression of FEM1B, PSMC2 and disulfide-isomerase A3 in MSCs treated with the liver differentiation protocol. These results from proteomic profiling will not only provide insight into the global responses of MSCs to hepatocyte differentiation, but will also lead to in-depth studies on the mechanisms of proteomic changes in MSCs.

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(A) The fat globules were demonstrated in the cytoplasm of MSCs cells treated with fat differentiation protocol; (B) Total RNA was isolated from MSCs cells treated with or without fat differentiation protocol. Expression of FEM1B, PSMC2 and disulfide-isomerase A3, mRNA levels was evaluated using RT-PCR. The results, based on the ratio of these mRNA amplification to that of GAPDH, are presented as the fold increase relative to the mRNA levels from MSC control. The results are expressed as the mean ± SD of three separate experiments (* p < 0.001 versus the control MSC).
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f5-ijms-11-04905: (A) The fat globules were demonstrated in the cytoplasm of MSCs cells treated with fat differentiation protocol; (B) Total RNA was isolated from MSCs cells treated with or without fat differentiation protocol. Expression of FEM1B, PSMC2 and disulfide-isomerase A3, mRNA levels was evaluated using RT-PCR. The results, based on the ratio of these mRNA amplification to that of GAPDH, are presented as the fold increase relative to the mRNA levels from MSC control. The results are expressed as the mean ± SD of three separate experiments (* p < 0.001 versus the control MSC).

Mentions: To study whether or not the expression of FEM1B, PSMC2 and disulfide-isomerase A3 was specific only for MSCs treated with the liver differentiation protocol, we measured the expression of these genes in the MSCs treated with fat differentiation protocol. These MSCs treated with fat differentiation protocol were derived from our previous study [6]. These cells demonstrated many fat globules in their cytoplasm (Figure 5A). The results demonstrated that the expression of disulfide-isomerase A3 was up-regulated in MSCs treated with fat differentiation protocol. However, there is no change in the expression of FEM1B and PSMC2 in MSCs treated with fat differentiation protocol when compared to the undifferentiated cells (Figure 5B).


Proteomic profiles of mesenchymal stem cells induced by a liver differentiation protocol.

Leelawat K, Narong S, Chaijan S, Sa-Ngiamsuntorn K, Disthabanchong S, Wongkajornsilp A, Hongeng S - Int J Mol Sci (2010)

(A) The fat globules were demonstrated in the cytoplasm of MSCs cells treated with fat differentiation protocol; (B) Total RNA was isolated from MSCs cells treated with or without fat differentiation protocol. Expression of FEM1B, PSMC2 and disulfide-isomerase A3, mRNA levels was evaluated using RT-PCR. The results, based on the ratio of these mRNA amplification to that of GAPDH, are presented as the fold increase relative to the mRNA levels from MSC control. The results are expressed as the mean ± SD of three separate experiments (* p < 0.001 versus the control MSC).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3100820&req=5

f5-ijms-11-04905: (A) The fat globules were demonstrated in the cytoplasm of MSCs cells treated with fat differentiation protocol; (B) Total RNA was isolated from MSCs cells treated with or without fat differentiation protocol. Expression of FEM1B, PSMC2 and disulfide-isomerase A3, mRNA levels was evaluated using RT-PCR. The results, based on the ratio of these mRNA amplification to that of GAPDH, are presented as the fold increase relative to the mRNA levels from MSC control. The results are expressed as the mean ± SD of three separate experiments (* p < 0.001 versus the control MSC).
Mentions: To study whether or not the expression of FEM1B, PSMC2 and disulfide-isomerase A3 was specific only for MSCs treated with the liver differentiation protocol, we measured the expression of these genes in the MSCs treated with fat differentiation protocol. These MSCs treated with fat differentiation protocol were derived from our previous study [6]. These cells demonstrated many fat globules in their cytoplasm (Figure 5A). The results demonstrated that the expression of disulfide-isomerase A3 was up-regulated in MSCs treated with fat differentiation protocol. However, there is no change in the expression of FEM1B and PSMC2 in MSCs treated with fat differentiation protocol when compared to the undifferentiated cells (Figure 5B).

Bottom Line: We demonstrated that MSCs treated with the liver differentiation protocol expressed significantly more albumin, CK19 and CK20, than did undifferentiated cells.In addition the results of proteomic study demonstrated increases expression of FEM1B, PSMC2 and disulfide-isomerase A3 in MSCs treated with the liver differentiation protocol.These results from proteomic profiling will not only provide insight into the global responses of MSCs to hepatocyte differentiation, but will also lead to in-depth studies on the mechanisms of proteomic changes in MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Rajavithi Hospital, Rajathevi, Bangkok, 10400, Thailand; E-Mails: kawin.leelawat@gmail.com (K.L.); sirilucknarong@hotmail.com (S.N.); bubu_b600@hotmail.com (S.C.).

ABSTRACT
The replacement of disease hepatocytes and the stimulation of endogenous or exogenous regeneration by human mesenchymal stem cells (MSCs) are promising candidates for liver-directed cell therapy. In this study, we isolated MSCs from adult bone marrow by plastic adhesion and induced differentiation with a liver differentiation protocol. Western blot analyses were used to assess the expression of liver-specific markers. Next, MSC-specific proteins were analyzed with two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). To confirm the results from the proteomic study, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analyses were performed. We demonstrated that MSCs treated with the liver differentiation protocol expressed significantly more albumin, CK19 and CK20, than did undifferentiated cells. In addition the results of proteomic study demonstrated increases expression of FEM1B, PSMC2 and disulfide-isomerase A3 in MSCs treated with the liver differentiation protocol. These results from proteomic profiling will not only provide insight into the global responses of MSCs to hepatocyte differentiation, but will also lead to in-depth studies on the mechanisms of proteomic changes in MSCs.

Show MeSH
Related in: MedlinePlus