Prenatal and postnatal transplantation of human amniotic fluid stem cells in spinal muscular atrophy mice

Abstract BackgroundType III spinal muscular atrophy (SMA) is a single gene disorder affecting motor function in uterus. Several types of stem cells were utilized to ameliorate SMA based on its capability of regeneration and differentiation. Amniotic fluid is an alternative source of stem cells and is safely sampled without ethical issues. Human amniotic fluid stem cell (hAFSC) shared common surface markers of mesenchymal stem cell. Therefore, this study aims to examine the therapeutic potential of hAFSC for SMA. MethodsOur SMA model mice were generated by deletion of exon 7 of Smn gene and knock-in of human SMN2. A total of 16 SMA model mice were injected with 1x105 hAFSC in uterus, and the other 16 mice served as the negative control. Motor function was analyzed by Rotarod maintenance test, tilting test and grasping test every two months. Twelve months after transplantation, all organs were extracted for post-mortem analysis. Engraftment of hAFSC in organs were assessed by flow cytometry and RNA scope. To observe the function of neuromuscular junction, frequency of myocytes, neurons and innervated receptors were estimated by H&E, methylene blue and immunocytochemistry staining. ResultsWith hAFSC transplantation, 15 fetuses from 5 dams survived (15 of 16, 93.75% survival) and showed better performance in all three motor function tests. Higher engraftment frequency in organs were observed in muscle and liver after hAFSC transplantation. Besides, the muscle of SMA mice with hAFSC transplantation expressed much laminin α and PAX-7. Significantly higher frequency of myocytes, neurons and innervated receptors were observed after hAFSC transplantation. ConclusionsIn our study, hAFSC engrafted on neuromuscular organs and improved cellular and behavioral outcomes of SMA model mice. This fetal therapy could preserve the time window and treat in the uterus to avoid irreversible damage.

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Amniotic Fluid Cells, Stem Cells, and p53: Can We Stereotype p53 Functions?

International Journal of Molecular Sciences ◽

10.3390/ijms20092236 ◽

2019 ◽

Vol 20 (9) ◽

pp. 2236 ◽

Cited By ~ 2

Author(s):

Melissa Rodrigues ◽

Christine Blattner ◽

Liborio Stuppia

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Adult Stem Cells ◽

Tumor Formation ◽

Human Stem Cells ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells ◽

Differentiated Cells ◽

High Proliferation Rate ◽

Alternative Sources

In recent years, great interest has been devoted to finding alternative sources for human stem cells which can be easily isolated, ideally without raising ethical objections. These stem cells should furthermore have a high proliferation rate and the ability to differentiate into all three germ layers. Amniotic fluid, ordinarily discarded as medical waste, is potentially such a novel source of stem cells, and these amniotic fluid derived stem cells are currently gaining a lot of attention. However, further information will be required about the properties of these cells before they can be used for therapeutic purposes. For example, the risk of tumor formation after cell transplantation needs to be explored. The tumor suppressor protein p53, well known for its activity in controlling Cell Prolif.eration and cell death in differentiated cells, has more recently been found to be also active in amniotic fluid stem cells. In this review, we summarize the major findings about human amniotic fluid stem cells since their discovery, followed by a brief overview of the important role played by p53 in embryonic and adult stem cells. In addition, we explore what is known about p53 in amniotic fluid stem cells to date, and emphasize the need to investigate its role, particularly in the context of cell tumorigenicity.

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291 ISOLATION AND CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM HUMAN AMNIOTIC FLUID

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab291 ◽

2011 ◽

Vol 23 (1) ◽

pp. 243 ◽

Cited By ~ 3

Author(s):

S.-A. Choi ◽

J.-H. Lee ◽

K.-J. Kim ◽

E.-Y. Kim ◽

K.-S. Park ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Amniotic Fluid ◽

Adult Stem Cells ◽

Embryonic Stem ◽

Stem Cell Marker ◽

Second Trimester ◽

Cell Marker ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

Adult stem cells have the capacity to differentiate into several different cell types, although their differentiation potential is limited compared with that of embryonic stem cells. Thus, adult stem cells are regarded as an exciting source for new cell therapies. Recent observations also indicate that stem cells derived from second-trimester amniocentesis are pluripotent – capable of differentiating into multiple lineages, including representatives of all 3 embryonic germ layers. In addition, amniotic fluid stem cells can be used in the generation of disease- or patient-specific stem cells, and amniotic fluid stem cells could be an ideal source for autologous cell replacement therapy in the later life of the fetus. The aim of the present study was to investigate isolation and characterisation of human amniotic fluid-derived mesenchymal stem cells (hAFS). We successfully isolated and characterised hAFS. Amniotic fluid samples were collected in the second trimester (median gestational age: 16 weeks, range: 15–17 weeks) for prenatal diagnosis. Specimens (2 mL) were centrifuged and incubated in low-glucose DMEM supplemented with 10% FBS, 25 ng of basic fibroblast growth factor, and 10 ng of epidermal growth factor at 37°C with 5% CO2. Human amniotic fluid cell (passage 6) expression of stem cell specific markers OCT-4, SOX2, Rex1, FGF4, and NANOG was confirmed by RT-PCR. Flow cytometric analysis showed that hAFS (passage 10) were positive for CD44, CD29, CD146, STRO1, and CD90 but negative for CD19. Immunocytochemical analysis of hAFS (passage 11) also showed the expression of OCT-4, SSEA-1, CD44, CD29, CD146, STRO1, and CD90, but hAFS were negative for CD19 and CD14. In conclusion, according to the previous studies on other mammalians, hAFS are an appropriate source of pluripotent stem cells. Here, we demonstrated that hAFS have a high expression of stem cell specific marker, including embryonic stem cell marker and mesenchymal stem cell marker. Therefore, amniotic fluid may be a suitable alternative source of multipotent stem cells.

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Renal tubular regeneration from human Amniotic Fluid Stem Cells

Journal of the American College of Surgeons ◽

10.1016/j.jamcollsurg.2005.06.100 ◽

2005 ◽

Vol 201 (3) ◽

pp. S47 ◽

Cited By ~ 1

Author(s):

Roger De Filippo ◽

Laura Perin ◽

Stefano Giuliani ◽

Akito Maeshima ◽

Daniel Jin ◽

...

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells ◽

Renal Tubular

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Human Amniotic Fluid Stem Cells: Therapeutic Potential for Perinatal Patients with Intractable Neurological Disease

The Keio Journal of Medicine ◽

10.2302/kjm.2017-0019-ir ◽

2018 ◽

Vol 67 (4) ◽

pp. 57-66 ◽

Cited By ~ 5

Author(s):

Daigo Ochiai ◽

Hirotaka Masuda ◽

Yushi Abe ◽

Toshimitsu Otani ◽

Marie Fukutake ◽

...

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Neurological Disease ◽

Therapeutic Potential ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

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Nuclear Nox4 Role in Stemness Power of Human Amniotic Fluid Stem Cells

Oxidative Medicine and Cellular Longevity ◽

10.1155/2015/101304 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 8

Author(s):

Tullia Maraldi ◽

Marianna Guida ◽

Manuela Zavatti ◽

Elisa Resca ◽

Laura Bertoni ◽

...

Keyword(s):

Stem Cells ◽

Transcription Factors ◽

Amniotic Fluid ◽

Human Stem Cells ◽

Human Amniotic Fluid ◽

Differentiation Potential ◽

Amniotic Fluid Stem Cells ◽

Target Molecules ◽

Cellular Compartments

Human amniotic fluid stem cells (AFSC) are an attractive source for cell therapy due to their multilineage differentiation potential and accessibility advantages. However the clinical application of human stem cells largely depends on their capacity to expandin vitro, since there is an extensive donor-to-donor heterogeneity. Reactive oxygen species (ROS) and cellular oxidative stress are involved in many physiological and pathophysiological processes of stem cells, including pluripotency, proliferation, differentiation, and stress resistance. The mode of action of ROS is also dependent on the localization of their target molecules. Thus, the modifications induced by ROS can be separated depending on the cellular compartments they affect. NAD(P)H oxidase family, particularly Nox4, has been known to produce ROS in the nucleus. In the present study we show that Nox4 nuclear expression (nNox4) depends on the donor and it correlates with the expression of transcription factors involved in stemness regulation, such as Oct4, SSEA-4, and Sox2. Moreover nNox4 is linked with the nuclear localization of redox sensitive transcription factors, as Nrf2 and NF-κB, and with the differentiation potential. Taken together, these results suggest that nNox4 regulation may have important effects in stem cell capability through modulation of transcription factors and DNA damage.

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