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Hwang, Kim, Jo, Lee, Koo, Song, Na, and Choi: Comparison with human amniotic membrane- and adipose tissue-derived mesenchymal stem cells
Original Article
Korean Journal of Obstetrics & Gynecology

Korean Journal of Obstetrics & Gynecology 2011; 54(11): 674-683.

Published online: 23 November 2011

DOI: https://doi.org/10.5468/KJOG.2011.54.11.674

Comparison with human amniotic membrane- and adipose tissue-derived mesenchymal stem cells

In Seob Hwang, MD, Hwi Gon Kim, MD, Deok Hyeon Jo, MD, Dong Hyung Lee, MD, Yun Hee Koo, MD, Yong Jung Song, MD, Yong Jin Na, MD, Ook Hwan Choi, MD

Department of Obstetrics and Gynecology, Pusan National University School of Medicine, Busan, Korea.

Corresponding author: Ook Hwan Choi, MD. Department of Obstetrics and Gynecology, Pusan National University Yangsan Hospital, Beomeo-ri, Mulgeum-eup, Yangsan 626-770, Korea. Tel: +82-55-360-2580, Fax: +82-55-360-2160, ohchoi@pusan.ac.kr

Received 22 March 2011       Revised 9 September 2011       Accepted 15 September 2011

Copyright © 2011. Korean Society of Obstetrics and Gynecology

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objective

Mesenchymal stem cells (MSC) have been considered as an ideal source of stem cells because of low immunogenicity and availability of autologous cells. Although it has been reported that amniotic membrane contains MSC, the difference between amniotic membrane-derived mesenchymal stem cells (AMSC) and MSC isolated from other tissues is still not clear. This study was designated to compare the characteristics and gene expression profi le of human AMSCs (hAMSC) and adipose tissue-derived mesenchymal stem cells (hADSC).

Methods

MSC were cultured from human amniotic membranes and adipose tissue by enzyme digestion. We compared the growth rate, surface marker expression, differentiation potential to adipogenic and osteogenic lineages, and gene expression of hAMSC with those of hADSC.

Results

hAMSC had growth rate and surface marker expression similar with hADSC. However, cyclopamine inhibited hAMSC proliferation in a dose-dependent manner without affecting hADSC proliferation. hAMSC showed lower differentiation potential to adipogenic and osteogenic lineages and lower in vivo tumor growth promoting effect in lung cancer cells xenotransplantion model of nude mouse than hADSC. Gene expression analysis using microarray revealed that many genes to be expressed differentially between hAMSC and hADSC are related to development and differentiation processes.

Conclusion

These findings indicate that hAMSC have different characteristics with hADSC, and that to use MSC isolated from different sources according to therapeutic purposes may provide more promising results in clinical trials than general use of MSC from a specific source.

Keywords: Mesenchymal stem cells, Human amniotic membrane-derived mesenchymal stem cells, Human adipose tissue-derived mesenchymal stem cells

Figures and Tables

Fig. 1
The cell morphology hAMSC and hADSC. Photographs were taken under phase microscope (×100). hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells.
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Fig. 2
(A) Proliferation of hAMSC and hADSC. (B) Effect of cyclopamine on the proliferation of hAMSC and hADSC. Cell number was determined 5 days after plating of cells in the presence or absence of cyclopamine. Data represent mean ± standard error of mean (SEM) of three different experiments. (C) Effect of cyclopamine on the viability of hAMSC and hADSC. Cell viability of hAMSC and hADSC in the absence or presence of cyclopamine was determined on days 4 by trypan blue exclusion assay. Data represent mean ± SEM of three different experiments. hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells. aP < 0.05 compared with the data in the absence of cyclopamine.
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Fig. 3
Adipogenic differentiation of hAMSC and hADSC. (A) Cells were grown to confl uence and then induced to osteogenic or adipogenic differentiation in differentiation media. Adipogenic differentiation (AM) was determined with Oil Red O staining as an indicator of intracellular lipid accumulation (×200). (B) The quantitation of adipogenic differentiation was performed by measurement of optical density in isopropanol extract of oil red O staining. Data represent mean ± SEM of three different experiments. hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells. aP < 0.05 compared with hADSC.
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Fig. 4
Osteogenic differentiation of hAMSC and hADSC. Cells were grown to confl uence and then induced to osteogenic differentiation in differentiation media. (A) Osteogenic differentiation (OM) was determined by calcification deposits on the cell monolayer, which were stained with alizarin red S Alizarin Red S. (×200). (B) The quantitation of osteogenic differentiation was performed by determination of density and area of Alizarin Red S staining with an image analysis program (Metamorph, Molecular Devices, LLC., Sunnyvale, CA, USA). Data represent mean ± standard error of mean of three different experiments. hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells. aP < 0.05 compared with hADSC.
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Fig. 5
Effects of hAMSC and hADSC injection on the tumor growth in vivo. (A) Effect of hASCs cotransplantation on tumor growth from xenotransplanted H460 cells. H460 (1×105) were injected subcutaneously and 1×106 hADSC or hAMSC were injected into left ventricle. Representative photograph of H460-derived tumors in nude mice on 14 days after transplantation. (B) Quantification of tumor weights. Data represent mean ± standard error of mean (n=4). hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells. aP < 0.05, significant difference from control data of H460 alone; bP < 0.05, significant difference from H460 + hADSC group.
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Table 1
Expression of surface markers in hADSC and hAMSC
kjog-54-674-i001

Data represent mean ± standard error of mean (n = 4 per group).

hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells.

Table 2
Genes list down-regulated in hAMSC than in hADSC
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hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells.

Table 3
Genes list up-regulated in hAMSC than hADSC
kjog-54-674-i003

hAMSC, human amniotic membrane-derived mesenchymal stem cells; hADSC, human adipose tissue-derived mesenchymal stem cells.

Table 4
Functional annotation clustering by DAVID bioinformatics
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DAVID, database for annotation, visualization and integrated discovery.

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