The adult human heart as a source for stem cells: repair strategies with embryonic-like progenitor cells

Abstract Bone marrow (BM) contains hematopoietic stem cells (HSCs), which can give rise to all mature blood cells and marrow stromal cells as well. Recently, it has been shown that non-hematopoietic stem/progenitor cells which can differentiate into non-hematopoietic tissues also reside in the BM. Although culture expanded cells have been studied in great detail, little is known about the phenotype and quantity of these cells in freshly harvested adult human BM. The aim of this study is to isolate and characterize hematopoietic and non-hematopoietic stem/progenitor cells in adult human BM by comparing two different isolation techniques and their effects on the yield of hematopoietic, mesenchymal and endothelial stem/progenitor cell populations. BM samples were collected mechanically from isolated rib specimens obtained during lung resection (n=10), or from BM aspirates harvested from the humerus of orthopedic patients (n=17). BM mononuclear cells were purified on a Ficoll/Hypaque density gradient and stained simultaneously using CD105 FITC, CD73 PE, CD34 ECD, CD90 PE.Cy5, CD117 PE.Cy7, CD133 APC, CD45 APC.Cy7 and DAPI as a marker of nucleated cells. 2–15 million cells per sample were acquired on a Dako CyAn cytometer and the data were analyzed offline using prototype analytical software (Venturi, Applied Cytometry Systems). The significant difference in the percentage of the CD45 − singlets (non-hematopoietic cells) between BM aspirates and rib-derived samples indicates hemodilution in the bone marrow aspirates. Although we have observed a slight difference in the mean of hematopoietic stem cell content between samples, it was not statistically significant. According to our results, the quantity of mesenchymal stem cells was higher in rib-derived BM than BM aspirates (p value=0.028). The expression of some stem/progenitor cell markers, such as CD90 (Thy-1), CD117 (c-Kit) and CD133 remained similar for all cell types. Our results are shown in the table below. Surface Antigens RibBM (n=10)¥ BMA (n=17)¥ p Value % % Total Cells CD45- of nucleated cells 15.3 ± 7.9 5.7 ± 5.2 0.004 CD34+ Hematopoietic Stem Cells (HSCs)* CD34 of CD45+ 1.7 ± 1.48 2.6 ± 2.0 0.883 CD117 74.6 ± 31.3 53.3 ± 18.8 0.073 CD90 60.3 ± 44.5 35.9 ± 36.5 0.134 CD133 70.3 ± 31.8 62.3 ± 21.4 0.443 Endothelial Progenitor Cells (EPCs)* EPCs of nucleated cells 0.05 ± 0.03 0.12 ± 0.2 0.323 CD117 81.3 ± 29.8 78.1 ± 20.2 0.746 CD90 66.7 ± 39.7 53.7 ± 31.4 0.356 CD133 45.9 ± 32.7 33.9 ± 22.0 0.265 Mesenchymal Stem Cells (MSCs)* MSCs of nucleated cells 0.086 ± 0.14 0.008 ± 0.01 0.028 CD117 60.2 ± 36.8 49.8 ± 34.3 0.471 CD90 66.0 ± 27.7 65.7 ± 29.1 0.981 CD133 37.8 ± 27.4 39.9 ± 28.9 0.857 RibBM: Rib-derived BM, BMA: Bone Marrow Aspirate ¥Data are given as mean ± SD. *CD90, CD117 and CD133 expressions are shown for each stem/progenitor fraction: Hematopoietic stem cells (CD34 + CD45 + and light scatter properties according to the ISHAGE protocol), endothelial progenitor cells (CD34bright CD45 − CD105 +) and mesenchymal stem cells (CD34 − CD45 − CD73 + CD105 +).

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European Society for Surgical Research: Biological Properties & Regenerative Potential, in Vitro & in Vivo, of Human Cardiac Stem Cells Isolated from the Adult Human Heart

Journal of Surgical Research ◽

10.1016/j.jss.2011.11.273 ◽

2012 ◽

Vol 172 (2) ◽

pp. 203

Author(s):

T. Theologou ◽

G.M. Ellison ◽

C. Vicinanza ◽

M. Torella ◽

V. Galuppo ◽

...

Keyword(s):

Stem Cells ◽

European Society ◽

Human Heart ◽

Biological Properties ◽

Surgical Research ◽

Cardiac Stem Cells ◽

Adult Human

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β1 - and β2 -adrenoceptor responses in cardiomyocytes derived from human embryonic stem cells: comparison with failing and non-failing adult human heart

British Journal of Pharmacology ◽

10.1038/sj.bjp.0707619 ◽

2008 ◽

Vol 153 (4) ◽

pp. 751-759 ◽

Cited By ~ 52

Author(s):

M Brito-Martins ◽

S E Harding ◽

N N Ali

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Human Heart ◽

Embryonic Stem ◽

Adult Human

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Human mesenchymal stem cells improve ex vivo expansion of adult human CD34+ peripheral blood progenitor cells and decrease their allostimulatory capacity

Experimental Hematology ◽

10.1016/j.exphem.2006.10.015 ◽

2007 ◽

Vol 35 (3) ◽

pp. 507-515 ◽

Cited By ~ 54

Author(s):

Na Li ◽

Pierre Feugier ◽

Brigitte Serrurrier ◽

Veronique Latger-Cannard ◽

Jean-François Lesesve ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Progenitor Cells ◽

Peripheral Blood ◽

Ex Vivo ◽

Human Mesenchymal Stem Cells ◽

Ex Vivo Expansion ◽

Adult Human ◽

Human Cd34 ◽

Peripheral Blood Progenitor Cells

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P2551The distribution and characteristics of endogenous cardiac stem cells in the atria and ventricle of the adult human heart

European Heart Journal ◽

10.1093/eurheartj/ehx502.p2551 ◽

2017 ◽

Vol 38 (suppl_1) ◽

Author(s):

S. Sarvananthan ◽

P.P. Punjabi ◽

F. Lewis ◽

N. Latif ◽

P. Sarathchandra ◽

...

Keyword(s):

Stem Cells ◽

Human Heart ◽

Cardiac Stem Cells ◽

Adult Human

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Abstract 1015: Human Heart Contains A Stem Cell Population With Mesenchymal Properties

Circulation ◽

10.1161/circ.116.suppl_16.ii_202-a ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Alessandra Rossini ◽

Alessandro Scopece ◽

Ombretta Pozzoli ◽

Giulio Pompilio ◽

Carlo Gaetano ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Population ◽

Human Heart ◽

Liver Fat ◽

Stem Cell Population ◽

Cardiac Markers ◽

Adult Human ◽

Hla Dr

Background Mesenchymal Stem Cells (MSC) were initially isolated from the bone marrow (BM) and identified as plastic adherent cells with a typical immunophenotype (negative for CD45, CD34, CD117 and HLA-DR and positive for CD105, CD90, CD44, CD73 and CD13), and able to differentiate into mesodermal lineages in vitro . Recently, MSC have been isolated from different adult tissues, i.e. liver, fat, kidney and thymus. Aim of the present study was to search for Cardiac Mesenchymal Stem Cells (C-MSC) in the adult human heart. Methods and Results : Auricle fragments were obtained from adult patients undergoing cardiac surgery. From these specimens we isolated a high proliferating plastic adherent cell population that, by FACS analysis, expressed mesenchymal markers (CD105, CD44, CD73, CD13 ) and was negative for hematopoietic markers (CD45, CD34) and HLA-DR. Further, in appropriate differentiating media, these cells exhibited osteogenic and adipogenic differentiation as cultures were positive for Oil Red O and Von Kossa staining used to mark intracellular lipid droplets and calcium salts deposits, respectively. Quantitative Real Time (qRT) -PCR analysis revealed that cells were negative for the early cardiac markers Nkx2.5 and Tbx5 and for the adult cardiac markers α-Myosin Heavy Chain and Myosin Light Chain-2a. Interestingly, in standard culture conditions, cells were positive for the early cardiac marker GATA4; in contrast, BM-MSC expressed GATA4 only after dexamethasone treatment. Additionally, when injected into the zebrafish blastula (about 100 –200 cells, n=70), C-MSC specifically homed in the cardiac region, as found at 24 –28 hpf (hours post fertilization). In contrast, transplanted BM-MSCs, were found not only in proximity of the heart, but throughout several mesodermal derivatives of the embryo (n=25). Conclusions: The adult human heart contains C-MSCs that can be easily expanded in vitro and may represent a useful population for autologous cell therapy of some cardiac diseases.

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