ACTA MEDICA LITUANICA. 2012. Vol. 19. No. 2. P . 75–79
© Lietuvos mokslų akademija, 2012
Surface markers distinguishing mesenchymal stem cells from fibroblasts
Gabrielis Kundrotas 1, 2
1 Biobank, Institute of Oncology, Vilnius University, Vilnius, Lithuania
2
Department of Botany and Genetics, Faculty of Natural Sciences, Vilnius University, Vilnius, Lithuania
Human mesenchymal stem cells (MSCs) are widely used for treatment of various diseases. Clinical applications require large quantities of MSCs, therefore these cells must be expanded in the culture system. It is believed that contamination of MSC cultures with fibroblasts may lead to the de-crease of the stem cell differentiation potential. Moreover, such stem cell preparations are potentially unsafe to use for clinical applications since a few fibroblasts can become tumorigenic. Therefore, there is a need to separate MSCs from fibroblasts. However, studies show that MSCs and fibroblasts have much in co
mmon. These two types of cells share such properties as identical spindle-like morphology, plastic adherence and the same expression of most surface antigens. The aim of this review ar-ticle is to analyze the literature on the similarities and differences between the MSCs and fibroblasts, particularly in the expression of cell surface markers in order to determine which could be used for quick separat-ing of MSCs from fibroblasts. Interestingly, the results of recent studies suggest that the use of CD10, CD26, CD106, CD146 and ITGA11 could be helpful for the discrimination of MSCs from fibroblasts. Identification and elimination of fibroblasts from MSC cultures could improve the MSC yield and differentiation potential and also prevent possible tumor forma-tion after MSC transplantation.
Key words: mesenchymal stem cells, fibroblasts, surface markers, senes-cence, tumorigenicity
Correspondence to: Gabrielis Kundrotas, Institute of Oncology, Vilnius University, Santariškių 1, LT-08660 Vilnius, Lithuania. E-mail: gabrielis.kundrotas@vuoi.lt
INTRODUCTION
Human mesenchymal stem cells (MSCs) due to their regenerative and immunomodulatory prop-erties are widely used for the treatment of bone and
cartilage damage, cardiovascular, gastrointestinal, autoimmune, neurodegenerative diseases and can-cer (1). It is shown that the use of MSCs in therapy is safe and can be effective (2). In 2006, the Inter-national Society for Cellular Therapy (ISCT) pro-posed the minimal criteria to define human MSCs. First, MSCs must be plastic-adherent when main-tained in the standard culture conditions. Second,
76Gabrielis Kundrotas
MSCs must express CD105, CD73 and CD90, and lack expression of CD45, CD34, CD14 or CD11b, CD79a or CD19 and HLA-DR surface molecules. Third, MSCs must differentiate into osteoblasts, adi p ocytes and chondroblasts in vitro (3). Fibro-blasts are terminally differentiated stromal cells (4) that provide mechanical strength to tissues by producing extracellular matrix and play a critical role during tissue development, differentiation and repair in many organs (5).
It is believed that contamination of MSC cul-tures with fibroblasts may lead to the decrease of the stem cell differentiation potential because fib-roblasts undergo senescence and eventually die. Moreover, such stem cell preparations are poten-tially unsafe to use for clinical applications since a few fibroblasts survive the crisis of senescence and can become tumorigenic (6). Therefore, there is
a need to separate MSCs from fibroblasts. Although MSCs and fibroblasts have been well studied, dif-ferences between these two cell types are not fully understood (7). MSCs and fibroblasts share much in common and the current definition suggested by the ISCT is thus incapable of separating MSCs from fibroblasts (8). The aim of this review article is to analyze the literature on the similarities and differences between the MSCs and fibroblasts, par-ticularly in the expression of cell surface markers in order to determine which could be used for quick separating of MSCs from fibroblasts. Similarities between mesenchymal stem cells and fibroblasts
Mesenchymal stem cells (MSCs) and fibroblasts exhibit a similar spindle-like morphology. In ad-dition to this, both types of cells adhere to plastic (7). Flow cytometry is a rapid method for sepa-ration of complex cell populations (9). However, MSCs and fibroblasts express the same surface markers. Alt E and colleagues (7) found that the expression of human adipose tissue-derived MSC surface markers CD44, CD90, CD105 was un-specific for these stem cells. Pure human embry-onic lung fibroblasts were also positive for these markers. Both hematopoietic cell markers (CD14, CD45) and the endothelial cell marker (CD31) were absent in MSCs and fibroblasts. Halfon and colleagues (10) reported coincidental results. It was shown that human bone marrow MSC (BM-MSC) surface markers CD9, CD29, CD44, CD73, CD90, CD105, CD166 were also expressed on human dermal fib
roblasts (10). Lorenz K and colleagues (11) showed similar expression pat-terns for CD14(–), CD29(+), CD31(–), CD34(–), CD44(+), CD45(–), CD71(+), CD73(+), CD90(+), CD105(+), CD133(–) and CD166(+) in human adipose tissue-derived stem cells and human der-mal skin-derived fibroblasts. Cappellesso-Fleury S and colleagues (4) compared the expression of 25 surface markers of human BM-MSCs and human dermal fibroblasts. They found the similar expres-sion patterns for 22 surface markers CD13(+), CD14(–), CD16(–), CD11a(–), CD33(–), CD34(–), CD43(–), CD45(–), CD49a(+), CD49b(+), CD54(+), CD86(–), CD90(+), CD105(+), CD117(–), CD146(+), CD164(+), CD166(+), CD138(variable), CD184(–), CD85k(–) and HLA-DR(–) in both types of cells.
Differences between mesenchymal stem cells and fibroblasts
永磁同步电动机
Despite the fact that in the studies reviewed in this article the majority of the investigated cell surface markers were nonspecific, CD106, CD146 and ITGA11 have been identified as MSC-spe-cific surface markers and CD10, CD26 as fibrob-last-specific surface markers (10). ITGA11 is a member of integrins that binds to collagen and is involved in cell attachment, cell migration and collagen reorganization on mesenchymal non-muscle cells (12). Halfon S and colleagues (10) showed that only 16.7% of fibroblasts expressed ITGA11 on their surface compared with MSCs of early (51.4%)
and late (28.6%) passages. CD106 (also known as vascular cell adhesion molecule-1 VCAM-1) is a member of the Ig superfamily which mediates leukocyte-endothelial cell adhe-sion and signal transduction during inflammation (13, 14). It was shown that CD106 protein was ex-pressed only on MSCs but not on fibroblasts (10). CD146 (also known as melanoma cell adhesion molecule MCAM) is important for endothelial cell migration and angiogenesis (15). It was shown that only 4.83% of fibroblasts were CD146 posi-tive compared with MSCs of early (91.7%) and late (79.8%) passages (10). Cappellesso-Fleury S and colleagues (4) reported different expression levels of CD10, CD106 and CD26 in comparison to 22 other markers. CD10 is a cell surface en-dopeptidase enzyme with neutral endopeptidase
activity and the ability to degrade a variety of bio-logically active compounds (16). CD26 is a cell surface glycoprotein known as dipeptidyl pepti-dase (DPP) IV and is involved in T lymphocyte activation (17). All fibroblasts were strongly posi-tive for CD26 and CD10 whereas less than 35% of BM-MSCs expressed CD10 (range: 16–35%) and CD26 expression was variable (range 40–78%). By contrast, more than 70% of BM-MSCs expressed CD106 whereas all fibroblasts were negative (4). All studies of the MSC and fibroblast surface marker expression reviewed in this article are sum
marized in Table.三星d500
Other important specific features of MSCs in which these cells differ from fibroblasts are the co-lony-forming capacity and differentiation potential (7). Actually, the results of investigations of the fi-broblast differentiation potential are controversial. Some authors state that fibroblasts do not differen-tiate into other types of cells (18). While the others show that fibroblasts do have the differentiation potential and even are potent immunoregulatory cells and functionally equivalent to mesenchymal stem cells (19). However, lately it has been sup-posed that such reported effects might be attribut-able to a great extent to the stem cell content within fibroblast preparations (7).
The need for separation of mesenchymal stem cells from fibroblasts
Clinical applications require large quantities of mesenchymal stem cells (MSCs). Expansion of cells in cultures is an attractive strategy because it makes it possible to administer more stem / pro-genitor cells than the patient can generate on his or her own. However, the differentiation potential of MSCs at later passages is often low. This phenome-non could be explained by contamination of MSC cultures with fibroblasts. It has been known for a long time that if fibroblasts from mouse embryos are cultured for prolonged periods, they undergo senescence followed by a “crisis” phase in
which many of the cells die. The few cells that survive the “crisis” first become immortal in the culture and then, after further expansion, can become tu-morigenic (6). Identification and elimination of fibroblasts from MSC cultures could improve the MSC yield and differentiation potential and also prevent tumor formation after MSC transplanta-tion (10).Table.Comparison of mesenchymal stem cell (MSC)
and fibroblast surface marker expression
CD9++(10)
CD10±+(4)
CD13++(4)
书录解题CD26Vari
able+(4)
CD29++(11), (10)
CD44++(7), (11),
(10)
CD49a++(4)
CD49b++(4)
CD54++(4)
CD71++(11)
CD73++(7), (11),
(10)
CD90++(4), (11),
(10)
CD105++(4), (7),
www.ddd20
(11), (10) CD106+–(4), (10)
CD138Vari-
able
Vari-
able(4)
CD146+±(4), (10) CD164++(4)
CD166++(4), (11),
(10) ITGA11+–(10)
CD11a––(4)
CD14
––(7), (11),
(4)
CD16––(4)
CD31––(7), (11) CD33––(4)
CD34––(4), (11) CD43––(4)
CD45
––(4), (7),
(11), (10) CD86––(4)
CD117––(4)
CD133––(11)
CD184––(4)
HLA-DR––(4) Note: +: expression; –: no or low expression.
78Gabrielis Kundrotas
CONCLUSIONS
It is believed that the contamination of mesenchy-mal stem cell (MSC) cultures with fibroblasts can be unsafe. However, MSCs and fibroblasts share much in common and the current definition of MSCs suggested in 2006 by the International So-ciety for Cellular Therapy is thus not capable of al-lowing us to discriminate between stem cells and fibroblasts any longer. Even then there were some thoughts that these criteria would probably require modification as new knowledge unfolded and that novel surface markers that might be identified in the future could lead to modifications of these cri-teria. The results of recent studies suggest that the use of CD10, CD26, CD106, CD146 and ITGA11 could be helpful for the discrimination of human bone marrow MSCs from human dermal fibro-blasts. However, there is a need to confirm these surface markers by investigating their expression on MSCs and fibroblasts isolated from other hu-man tissues. Eventually, such markers could be used for the quality control of MSC cultures after expansion, cryopreservation, gene transfection and other
manipulations.
Received 29 March 2012
Accepted 18 June 2012 References
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MEZENCHIMINES KAMIENINES LĄSTELES NUO FIBROBLASTŲ ATSKIRIANTYS
PAVIRŠIAUS ŽYMENYS
Santrauka
Žmogaus mezenchiminės kamieninės ląstelės (MKL) plačiai naudojamos įvairioms ligoms gydyti. Klinikinėje praktikoje reikalingi dideli MKL kiekiai, todėl šios ląste-lės turi būti padaugintos kultūrose. Manoma, kad MKL kultūrose esantys fibroblastai yra susiję su silpnėjančiu MKL diferenciacijos potencialu. Tokius kamieninių ląs-telių preparatus pavojinga naudoti terapijoje, nes dalis fibroblastų g
ali supiktybėti, todėl fibroblastus iš MKL kultūrų reikia pašalinti. Mokslinių tyrimų rezultatai rodo, kad MKL ir fibroblastai yra labai panašūs. Šioms abiejų tipų ląstelėms būdinga identiška verpstės formos morfologija, adhezija prie plastikinių paviršių ir panaši daugumos paviršiaus žymenų raiška. Šio straipsnio tiks-las – apžvelgti literatūrą apie MKL ir fibroblastų panašu-mus ir skirtumus, daugiausia dėmesio skiriant tyrimams, susijusiems su ląstelių paviršiaus žymenų, pagal kuriuos būtų galima greitai atskirti MKL nuo fibroblastų, raiška. Fibroblastų atpažinimas ir pašalinimas iš MKL kultūrų turėtų lemti didesnius auginamų MKL kiekius, stipres-nį šių kamieninių ląstelių diferenciacijos potencialą ir padėtų išvengti galimo navikų formavimosi po MKL transplantacijos. Naujų tyrimų rezultatai rodo, kad ląs-telių paviršiaus žymenys CD10, CD26, CD106, CD146 ir ITGA11 gali būti naudingi siekiant atskirti MKL nuo fibroblastų.
Raktažodžiai:mezenchiminės kamieninės ląstelės, fibroblastai, paviršiaus žymenys, senėjimas, navikų for-mavimasis
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