Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Sep;23(9):5981-5993.
doi: 10.1111/jcmm.14456. Epub 2019 Jul 9.

Effect of neuron-derived neurotrophic factor on rejuvenation of human adipose-derived stem cells for cardiac repair after myocardial infarction

Kun Yang  1   2 , Hui-Fang Song  1   3 , Sheng He  1   4 , Wen-Juan Yin  1 , Xue-Mei Fan  1   2 , Feng Ru  5 , Hui Gong  1 , Xiao-Yan Zhai  6 , Jie Zhang  1 , Ze-Xu Peng  1 , Guang-Xia Xi  2 , Jun Xie  1 , Ren-Ke Li  7   8
Affiliations

Effect of neuron-derived neurotrophic factor on rejuvenation of human adipose-derived stem cells for cardiac repair after myocardial infarction

Kun Yang et al. J Cell Mol Med. 2019 Sep.

Abstract

The decline of cell function caused by ageing directly impacts the therapeutic effects of autologous stem cell transplantation for heart repair. The aim of this study was to investigate whether overexpression of neuron-derived neurotrophic factor (NDNF) can rejuvenate the adipose-derived stem cells in the elderly and such rejuvenated stem cells can be used for cardiac repair. Human adipose-derived stem cells (hADSCs) were obtained from donors age ranged from 17 to 92 years old. The effects of age on the biological characteristics of hADSCs and the expression of ageing-related genes were investigated. The effects of transplantation of NDNF over-expression stem cells on heart repair after myocardial infarction (MI) in adult mice were investigated. The proliferation, migration, adipogenic and osteogenic differentiation of hADSCs inversely correlated with age. The mRNA and protein levels of NDNF were significantly decreased in old (>60 years old) compared to young hADSCs (<40 years old). Overexpression of NDNF in old hADSCs significantly improved their proliferation and migration capacity in vitro. Transplantation of NDNF-overexpressing old hADSCs preserved cardiac function through promoting angiogenesis on MI mice. NDNF rejuvenated the cellular function of aged hADSCs. Implantation of NDNF-rejuvenated hADSCs improved angiogenesis and cardiac function in infarcted mouse hearts.

Keywords: NDNF; ageing; cardiac injury repair; human adipose-derived stem cells; rejuvenation.

PubMed Disclaimer

Conflict of interest statement

The authors confirm that there are no conflicts of interest.

Figures

Figure 1
Figure 1
Cultivation and identification of hADSCs. A, General morphological observation of human adipose‐derived stem cells (hADSCs) cultured for 7 days after isolation from donors (17‐92 years old). B, Rate of cell growth was significantly higher in young than old hADSCs, n = 6/group, **P < 0.01 Young vs Old. C, Representative histogram plots of cell surface markers from young and old hADSCs. D, Comparison of % positive cells of hADSCs from young and old donors. n = 3/group
Figure 2
Figure 2
The proliferation, migration and differentiation of hADSCs decreased with age. A, Representative micrographs of immunofluorescent staining for 5‐bromo‐2'‐deoxyuridine (BrdU, red) with nuclei stained blue with DAPI. The percentage of BrdU+ cells decreased with age. B, Representative images showed the cell migration of human adipose‐derived stem cells (hADSCs) using the wound‐healing cell migration assay. Migration rate decreased with age. C, Representative micrographs of adipogenic differentiation stained for oil red ‘O’. Mature adipose cells were stained in red in both old and young groups. The adipogenic differentiation rate decreased with age. D, Representative micrographs of osteogenic differentiation stained for alizarin red. The osteocytes were stained in red in both old and young groups. The osteogenic differentiation rate decreased with age
Figure 3
Figure 3
NDNF expression decreased with age. A, The cell regeneration‐related genes, Sirt1 (sirtuin), Sirt2, Sirt6, Bmi1 (Polycomb complex protein BMI‐1), and Cbx8 (chromobox homolog 8), in human adipose‐derived stem cells (hADSCs) negatively correlated with age. B, The cell senescence‐related gene P16 (cyclin‐dependent kinase inhibitor 2A) increased with age. C, Neuron‐derived neurotrophic factor (NDNF) mRNA expression showed inverse correlation with age. The mRNA (D, n = 5/group) and protein (E, n = 3/group) levels of NDNF in the elderly group was significantly lower than that of the youth group. *P < 0.05, **P < 0.01
Figure 4
Figure 4
NDNF transduction rejuvenated old hADSCs by increasing proliferation and migration. A, Old human adipose‐derived stem cells (hADSCs) were transduced with a lentiviral vector overexpressing neuron‐derived neurotrophic factor (NDNF) which was also tagged with green fluorescent protein (GFP). Representative micrographs showed the GFP+ cells to indicate the transduction efficiency. There was no difference in transduction efficiency between old hADSCs transduced with NDNF (Old + NDNF) or with empty viruses (Old), n = 6/group. B, NDNF mRNA expression evaluated by RT‐PCR was significantly higher in Old + NDNF compared to Old group, n = 3/group. C, NDNF protein expression evaluated by Western blotting was significantly higher in Old + NDNF compared with that of the Old group, n = 3/group. D, Representative micrographs of immunofluorescent staining for 5‐bromo‐2'‐deoxyuridine (BrdU in red) with nuclei stained blue with DAPI. The percentage of BrdU+ cells was significantly higher in Old + NDNF compared with that of the Old group, n = 6/group. E, Representative images showed cell migration of hADSCs using the wound‐healing cell migration assay. Migration rate was significantly higher in Old + NDNF compared with empty vector‐transduced hADSCs (Old), n = 6/group. *P < 0.05, **P < 0.01
Figure 5
Figure 5
In vivo implantation of NDNF‐overexpressing old hADSCs improved cardiac function after MI. A, Representative M‐mode echocardiographic images taken before (baseline), and 7 and 28 days after myocardial infarction (MI) in mice that received implantation of control medium (Medium), empty vector‐transduced old human adipose‐derived stem cells (hADSCs, Old), neuron‐derived neurotrophic factor (NDNF)‐transduced old hADSCs (Old + NDNF) and untransduced young hADSCs (Young). The left ventricular fractional shortening (B, FS %), ejection fraction (C, EF %), left ventricular internal dimension‐diastole (D, LVIDd) and left ventricular internal dimension in systole (E, LVIDs ), of mice before and 7, 14, 21, and 28 days after MI, reflecting the changes in cardiac function of mice. FS and EF were significantly higher whereas LVIDs and LVIDd were significantly lower in the Young and Old + NDNF compared to the Old and Medium groups at 28 days after MI. n = 7/group, *P < 0.05,**P < 0.01
Figure 6
Figure 6
In vivo implantation of NDNF‐overexpressing old hADSCs decreased scar size and increased scar thickness in infarcted mouse hearts. A, Representative heart sections 28 days after myocardial infarction (MI) in mice that received implantation of control medium (Medium), empty vector‐transduced old human adipose‐derived stem cells (hADSCs, Old), neuron‐derived neurotrophic factor (NDNF)‐transduced old hADSCs (Old + NDNF) and untransduced young hADSCs (Young). B, Representative heart slides stained with Masson's trichrome and planimetry‐based quantification revealed that the scar size area was larger in the Medium and Old groups than in the Old + NDNF and Young groups at 28 days after MI (C). D, The scar thickness was greater in the Old + NDNF and Young groups than in the Medium and Old groups at 28 days after MI, n = 6/group
Figure 7
Figure 7
In vivo implantation of NDNF‐overexpressing old hADSCs increased angiogenesis and arteriole genesis. A, Western blotting analysis of the expression of NDNF protein in the infarcted area of mouse hearts that received implantation of control medium (Medium), empty vector‐transduced old human adipose‐derived stem cells (hADSCs, Old), neuron‐derived neurotrophic factor (NDNF)‐transduced old hADSCs (Old + NDNF) and untransduced young hADSCs (Young). B, NDNF protein levels were significantly higher in the Old + NDNF and Young groups compared with the Old and Medium control groups, n = 3/group. C, Representative micrographs of immunofluorescent staining for von Willebrand factor (vWF, red) with nuclei stained blue with DAPI. D, Capillary density was significantly higher in the Old + NDNF and Young groups compared with the Old and Medium control groups, n = 6/group. E, Representative micrographs of immunofluorescent staining for α‐smooth muscle actin (α‐SMA, green) with nuclei stained blue with DAPI. F, Arteriole density was significantly higher in the Old + NDNF and Young groups compared with the Old and Medium control groups, n = 6/group
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Arslan F, Bongartz L, ten Berg JM, et al. 2017 ESC guidelines for the management of acute myocardial infarction in patients presenting with ST‐segment elevation: comments from the Dutch ACS working group. Neth Heart J. 2018;26:417‐421. - PMC - PubMed
    1. Moran AE, Forouzanfar MH, Roth GA, et al. The global burden of ischemic heart disease in 1990 and 2010: the Global Burden of Disease 2010 study. Circulation. 2014;129:1493‐1501. - PMC - PubMed
    1. Mendis S, Davis S, Norrving B. Organizational update: the world health organization global status report on noncommunicable diseases 2014; one more landmark step in the combat against stroke and vascular disease. Stroke. 2015;46:e121‐e122. - PubMed
    1. Task Force on the management of STseamiotESoC , Steg PG, James SK, et al. Guidelines for the management of acute myocardial infarction in patients presenting with ST‐segment elevation. Eur Heart J. 2012;33:2569‐2619. - PubMed
    1. Reinsch M, Weinberger F. Stem cell‐based cardiac regeneration after myocardial infarction. Herz. 2018;43:109‐114. - PubMed

Publication types

MeSH terms

Substances