Prenatal Transplantation of Human Amniotic Fluid Stem Cell Could Improve Clinical Outcome of Type III Spinal Muscular Atrophy

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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Prenatal transplantation of human amniotic fluid stem cell could improve clinical outcome of type III spinal muscular atrophy in mice

Scientific Reports ◽

10.1038/s41598-021-88559-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Steven W. Shaw ◽

Shao-Yu Peng ◽

Ching-Chung Liang ◽

Tzu-Yi Lin ◽

Po-Jen Cheng ◽

...

Keyword(s):

Stem Cell ◽

Amniotic Fluid ◽

Spinal Muscular Atrophy ◽

Motor Function ◽

Time Window ◽

Therapeutic Potential ◽

Muscular Atrophy ◽

Single Gene ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cell

AbstractSpinal muscular atrophy (SMA) is a single gene disorder affecting motor function in uterus. Amniotic fluid is an alternative source of stem cell to ameliorate SMA. Therefore, this study aims to examine the therapeutic potential of Human amniotic fluid stem cell (hAFSC) for SMA. Our 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 1 × 105 hAFSC in uterus, and the other 16 mice served as the negative control. Motor function was analyzed by three behavioral tests. Engraftment of hAFSC in organs were assessed by flow cytometry and RNA scope. Frequency of myocytes, neurons and innervated receptors were estimated by staining. With hAFSC transplantation, 15 fetuses survived (93.75% survival) and showed better performance in all motor function tests. Higher engraftment frequency were observed in muscle and liver. Besides, the muscle with hAFSC transplantation expressed much laminin α and PAX-7. Significantly higher frequency of myocytes, neurons and innervated receptors were observed. In 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.

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

Cytotherapy ◽

10.1016/j.jcyt.2015.03.598 ◽

2015 ◽

Vol 17 (6) ◽

pp. S83

Author(s):

Steven Shaw ◽

Li-Kai Tsai ◽

Po-Jen Cheng

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Spinal Muscular Atrophy ◽

Muscular Atrophy ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

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Prenatal transplantation of human amniotic fluid stem cells for spinal muscular atrophy

Current Opinion in Obstetrics & Gynecology ◽

10.1097/gco.0000000000000444 ◽

2018 ◽

Vol 30 (2) ◽

pp. 111-115 ◽

Cited By ~ 2

Author(s):

Shao-Yu Peng ◽

Sheng-Wen S. Shaw

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Spinal Muscular Atrophy ◽

Muscular Atrophy ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

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Human Amniotic Fluid Mesenchymal Stem Cells from Second- and Third-Trimester Amniocentesis: Differentiation Potential, Molecular Signature, and Proteome Analysis

Stem Cells International ◽

10.1155/2015/319238 ◽

2015 ◽

Vol 2015 ◽

pp. 1-15 ◽

Cited By ~ 36

Author(s):

Jurate Savickiene ◽

Grazina Treigyte ◽

Sandra Baronaite ◽

Giedre Valiuliene ◽

Algirdas Kaupinis ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Amniotic Fluid ◽

Expression Patterns ◽

Specific Gene ◽

Specific Cell ◽

Third Trimester ◽

Human Amniotic Fluid ◽

Differentiation Potential

Human amniotic fluid stem cells have become an attractive stem cell source for potential applications in regenerative medicine and tissue engineering. The aim of this study was to characterize amniotic fluid-derived mesenchymal stem cells (AF-MSCs) from second- and third-trimester of gestation. Using two-stage protocol, MSCs were successfully cultured and exhibited typical stem cell morphological, specific cell surface, and pluripotency markers characteristics. AF-MSCs differentiated into adipocytes, osteocytes, chondrocytes, myocytes, and neuronal cells, as determined by morphological changes, cell staining, and RT-qPCR showing the tissue-specific gene presence for differentiated cell lineages. Using SYNAPT G2 High Definition Mass Spectrometry technique approach, we performed for the first time the comparative proteomic analysis between undifferentiated AF-MSCs from late trimester of gestation and differentiated into myogenic, adipogenic, osteogenic, and neurogenic lineages. The analysis of the functional and expression patterns of 250 high abundance proteins selected from more than 1400 demonstrated the similar proteome of cultured and differentiated AF-MSCs but the unique changes in their expression profile during cell differentiation that may help the identification of key markers in differentiated cells. Our results provide evidence that human amniotic fluid of second- and third-trimester contains stem cells with multilineage potential and may be attractive source for clinical applications.

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Nusinersen in later-onset spinal muscular atrophy

Neurology ◽

10.1212/wnl.0000000000007527 ◽

2019 ◽

Vol 92 (21) ◽

pp. e2492-e2506 ◽

Cited By ~ 55

Author(s):

Basil T. Darras ◽

Claudia A. Chiriboga ◽

Susan T. Iannaccone ◽

Kathryn J. Swoboda ◽

Jacqueline Montes ◽

...

Keyword(s):

Spinal Muscular Atrophy ◽

Motor Function ◽

Muscular Atrophy ◽

Compound Muscle Action Potential ◽

Extension Study ◽

Type Ii ◽

Open Label ◽

Type Iii ◽

Link Type ◽

Phase 1B

ObjectiveTo report results of intrathecal nusinersen in children with later-onset spinal muscular atrophy (SMA).MethodsAnalyses included children from a phase 1b/2a study (ISIS-396443-CS2; NCT01703988) who first received nusinersen during that study and were eligible to continue treatment in the extension study (ISIS-396443-CS12; NCT02052791). The phase 1b/2a study was a 253-day, ascending dose (3, 6, 9, 12 mg), multiple-dose, open-label, multicenter study that enrolled children with SMA aged 2–15 years. The extension study was a 715-day, single-dose level (12 mg) study. Time between studies varied by participant (196–413 days). Assessments included the Hammersmith Functional Motor Scale–Expanded (HFMSE), Upper Limb Module (ULM), 6-Minute Walk Test (6MWT), compound muscle action potential (CMAP), and quantitative multipoint incremental motor unit number estimation. Safety also was assessed.ResultsTwenty-eight children were included (SMA type II, n = 11; SMA type III, n = 17). Mean HFMSE scores, ULM scores, and 6MWT distances improved by the day 1,150 visit (HFMSE: SMA type II, +10.8 points; SMA type III, +1.8 points; ULM: SMA type II, +4.0 points; 6MWT: SMA type III, +92.0 meters). Mean CMAP values remained relatively stable. No children discontinued treatment due to adverse events.ConclusionsNusinersen treatment over ∼3 years resulted in motor function improvements and disease activity stabilization not observed in natural history cohorts. These results document the long-term benefit of nusinersen in later-onset SMA, including SMA type III.Clinicaltrials.gov identifierNCT01703988 (ISIS-396443-CS2); NCT02052791 (ISIS-396443-CS12).Classification of evidenceThis study provides Class IV evidence that nusinersen improves motor function in children with later-onset SMA.

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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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