Korean J Hepatobiliary Pancreat Surg.  2013 May;17(2):53-59. 10.14701/kjhbps.2013.17.2.53.

Hepatogenic differentiation of human mesenchymal stem cells from peritoneal adipose tissue

• Hong TH
• Park IY

• KMID: 1429979
• DOI: http://doi.org/10.14701/kjhbps.2013.17.2.53

Abstract

BACKGROUNDS/AIMS
It has been reported that functional hepatogenic differentiation has the possibility to occur in subcutaneous adipose tissue-derived stem cells. However, no studies have investigated whether the adipose tissue-driven stem cells present in various body parts differ according to hepatogenic differentiations. In this study, stem cells were separated from body visceral fat and abdominal subcutaneous adipose tissue, and cultured, and then hepatogenic differentiation was induced. We aim to investigate the possibilities and aspects of hepatogenic differentiations within the two types of fat cells.
METHODS
Omental fat tissues were obtained as visceral fat and abdominal subcutaneous adipose tissues were obtained from patients who had suction-assisted lipectomy. Stem cells were separated from the obtained fat tissues, and then, hepatogenic differentiation was carried out by utilizing 2-step differentiation protocols.
RESULTS
After the differentiation, two types of cultured cells that showed the similar neuron-like shapes were changed to cuboidal shapes and included several binucleated cells which could be characteristics of mature hepatocytes. We confirmed that hepatocyte specific genes and proteins such as albumin and CYP3A4 were being expressed. By utilizing the ELISA test, we were able to observe that the albumin was secreted into the culture fluids in both cells. After completing the differentiation, we observed the presence of the hepatocyte specific properties by confirming glycogen storage within the cells and the ICG reagent uptake.
CONCLUSIONS
We confirmed that hepatogenic differentiation was possible to occur in the omental fat as well as subcutaneous adipose tissue.

Keyword

Subcutaneous adipose tissue; Omental fat; Stem cell; Hepatogenic differentiation

MeSH Terms

Adipose Tissue
Cells, Cultured
Enzyme-Linked Immunosorbent Assay
Glycogen
Hepatocytes
Human Body
Humans
Intra-Abdominal Fat
Lipectomy
Mesenchymal Stromal Cells
Proteins
Stem Cells
Subcutaneous Fat
Subcutaneous Fat, Abdominal
Glycogen
Proteins

Figure

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  Fig. 1 Morphology of MSC from abdominal subcutaneous adipose tissue and omental fat during the differentiation protocol. Before differentiation culture, MSC from abdominal subcutaneous adipose tissue (A) and omental fat (B) showed bipolar neuron-like morphology. However, both types of MSC from abdominal subcutaneous adipose tissue (C) and omental fat (D) significantly changed the morphology, and developed a cuboidal shape and many of them appeared to be binucleated cells (arrows), which are typical hepatocytes after differentiation steps (magnification: A, B ×100; C, D ×200).

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  Fig. 2 RT-PCR analysis of the expressions of mRNA for ALB and CYP3A4 (compared with that of the human hepatoma cells, Hep G2). The expression of ALB in MSC from abdominal subcutaneous adipose tissue (black bars) and in MSC from omental fat (white bars) were detected at all time points and increased with differentiation time (A). The expression of CYP3A4 was detected at 2 weeks post-induction and then increased with differentiation time (B).

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  Fig. 3 Albumin secretion by both types of ADSC from abdominal subcutaneous adipose tissue (A) and omental fat (B). The amount of albumin released into the medium before and after hepatic differentiation in vitro analyzed by ELISA is shown. Data are expressed as mean±SEM of three independent experiments.

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  Fig. 4 PAS stainings for glycogen storage. After differentiation steps, MSC from omental fat (B) were strongly stained with purple color in cytoplasm, while undifferentiated MSC (A) were weakly stained (magnification: A, B ×200).

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  Fig. 5 Indocyanine green (ICG) cellular uptake. Undifferentiated MSC from omental fat (A) showed little stainings after ICG assays, whereas differentiated MSC (B) exhibited intense green stainings (magnification: A, B ×200).

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