Isolation, proliferation, cytogenetic, and molecular characterization and in vitro differentiation potency of canine stem cells from foetal adnexa: A comparative study of amniotic fluid, amnion, and umbilical cord matrix

2011 ◽  
Vol 78 (5) ◽  
pp. 361-373 ◽  
Author(s):  
M. Filioli Uranio ◽  
L. Valentini ◽  
A. Lange-Consiglio ◽  
M. Caira ◽  
A. C. Guaricci ◽  
...  
Keyword(s):  
Stem Cells ◽  
Comparative Study ◽  
Amniotic Fluid ◽  
Molecular Characterization ◽  
Umbilical Cord ◽  
In Vitro Differentiation ◽  
Umbilical Cord Matrix

312 ISOLATION, PROLIFERATION, AND CHARACTERIZATION OF MESENCHYMAL STEM CELLS FROM AMNIOTIC FLUID, AMNION, AND UMBILICAL CORD MATRIX IN THE DOG

2011 ◽  
Vol 23 (1) ◽  
pp. 252 ◽  
Author(s):  
L. Valentini ◽  
M. Filioli Uranio ◽  
A. Lange Consiglio ◽  
A. C. Guaricci ◽  
M. Caira ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Cell Viability ◽  
Umbilical Cord ◽  
Cell Biology ◽  
Cell Types ◽  
Telomerase Activity ◽  
Umbilical Cord Matrix

Mesenchymal stem cells (MSC) are defined as multipotent stem cells that can differentiate into various cell types in vivo and in vitro under controlled conditions. These cells express specific markers detectable by analysis at the mRNA or protein level. Important sources of MSC could be fetal adnexa, such as amniotic fluid (AF), amnion (AM), and umbilical cord matrix (UCM). Canine MSC should be of use for cell-based therapies and tissue engineering improving treatment of several diseases. Moreover, the dog has been considered an attractive animal model to study human diseases. In the present study, we successfully isolated and molecularly characterised AF-MSC, AM-MSC, and UCM-MSC from dogs. Chromosomal stability and telomerase activity were also investigated. Samples were recovered after elective ovariohysterectomy in 3 bitches 25 to 40 days of gestational age. After isolation, cells were maintained in culture (Bossolasco et al. 2006 Cell Res. 16, 329–336) for different passages to perform growth and doubling time (DT) studies. Expression analyses of embryonic (Oct-4, Nanog), mesenchymal (CD44, CD184, CD29), and haematopoietic (CD34, CD45) markers were carried out by RT-PCR. Karyotype analysis was performed by Q banding. Telomerase activity was analysed by TRAPeze Telomerase Detection Kit. In all 3 cell types, the morphology of proliferating cells appeared typically fibroblast-like. In the growth study, cells isolated from AF and AM were cultured until P3, and cells isolated from UCM were maintained until P7. The population DT in AF-MSC was significantly increased (Student’s t-test: P < 0.05) when comparing P1 v. P4. In AM-MSC, DT increased significantly in P1 v. P2 (P < 0.001), and in UCM-MSC, DT significantly increased in P1 v. P4 (P < 0.001). In AF-MSC, cell viability did not change with passages. In AM-MSC, cell viability significantly decreased (P < 0.001) between P1 and P4. In UCM-MSC, cell viability remained at approximately constant levels up to P6 and significantly decreased at P7 (P < 0.001). Amnion and UCM-MSC expressed Oct-4 and CD44, CD184, and CD29, whereas AF-MSC expressed only Oct-4 and CD44. Nanog, CD34, and CD45 were never found to be expressed in any cell line at any passage. In all cell lines, analysed metaphases at P4 showed normal chromosomal number and structure. Telomerase activity was observed in UCM-MSC, whereas tests on AF and AM-MSC are still on going. We first reported data on isolation, in vitro culture, and characterisation of MSC from AM and UCM in the dog. Cells expressed embryonic and MSC markers beginning at P1 and showed normal karyotype. These data indicated that canine MSC from fetal adnexa could be used to study stem cell biology and their application in therapeutic programs. Financial support was provided by Fondi di Ateneo 2009. University of Bari Aldo Moro (COD. ORBA09UDWX) (Resp. Sci. Maria Elena Dell’Aquila).

scholarly journals In vitro differentiation of human oocyte-like cells from oogonial stem cells: single-cell isolation and molecular characterization

2018 ◽  
Vol 33 (3) ◽  
pp. 464-473 ◽  
Author(s):  
Erica Silvestris ◽  
Paola Cafforio ◽  
Stella D’Oronzo ◽  
Claudia Felici ◽  
Franco Silvestris ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Molecular Characterization ◽  
Cell Isolation ◽  
Human Oocyte ◽  
In Vitro Differentiation ◽  
Single Cell Isolation

In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp

2013 ◽  
Vol 45 (8) ◽  
pp. 669-676 ◽  
Author(s):  
Gianluca Carnevale ◽  
Massimo Riccio ◽  
Alessandra Pisciotta ◽  
Francesca Beretti ◽  
Tullia Maraldi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
Dental Pulp ◽  
In Vitro Differentiation ◽  
Β Cells ◽  
Human Amniotic Fluid

scholarly journals Insulin Promotes the Proliferation of Human Umbilical Cord Matrix-Derived Mesenchymal Stem Cells by Activating the Akt-Cyclin D1 Axis

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Peng Li ◽  
Jinsong Wei ◽  
Xiang Gao ◽  
Bo Wei ◽  
Hao Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cyclin D1 ◽  
Umbilical Cord ◽  
Cell Cycle Progression ◽  
Human Umbilical Cord ◽  
Expression Levels ◽  
Umbilical Cord Matrix ◽  
Phosphorylated Akt

Background. The functions of insulin in mesenchymal stem cells (MSC) remain poorly understood. Methods. MSC from human umbilical cord matrix (UCM) cultured in serum-free media (SFM) with or without insulin were subjected to various molecular biological analyses to determine their proliferation and growth states, expression levels of Akt-cyclin D1 signaling molecules, and in vitro differentiation capacities. Results. Insulin accelerated the G1-S cell cycle progression of UCM-MSC and significantly stimulated their proliferation and growth in SFM. The pro-proliferative action of insulin was associated with augmented cyclin D1 and phosphorylated Akt expression levels. Akt inactivation remarkably abrogated insulin-induced increases in cyclin D1 expression and cell proliferation, indicating that insulin enhances the proliferation of UCM-MSC via acceleration of the G1-S transition mediated by the Akt-cyclin D1 pathway. Additionally, the UCM-MSC propagated in SFM supplemented with insulin exhibited similar specific surface antigen profiles and differentiation capacities as those generated in conventional media containing fetal bovine serum. Conclusions. These findings suggest that insulin acts solely to promote UCM-MSC proliferation without affecting their immunophenotype and differentiation potentials and thus have important implications for utilizing insulin to expand clinical-grade MSC in vitro.

The In Vitro Differentiation of GDNF Gene-Engineered Amniotic Fluid-Derived Stem Cells into Renal Tubular Epithelial-Like Cells

2018 ◽  
Vol 27 (9) ◽  
pp. 590-599 ◽  
Author(s):  
Ying Lu ◽  
Zhuojun Wang ◽  
Lu Chen ◽  
Jia Wang ◽  
Shulin Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
In Vitro Differentiation ◽  
Gdnf Gene ◽  
Renal Tubular

Characterization and in vitro differentiation potency of early-passage canine amnion- and umbilical cord-derived mesenchymal stem cells as related to gestational age

2014 ◽  
Vol 81 (6) ◽  
pp. 539-551 ◽  
Author(s):  
Manuel Filioli Uranio ◽  
Maria Elena Dell'Aquila ◽  
Michele Caira ◽  
Antonio Ciro Guaricci ◽  
Mario Ventura ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Gestational Age ◽  
In Vitro Differentiation ◽  
Early Passage

scholarly journals In vitro differentiation of human umbilical cord blood mesenchymal stem cells into functioning hepatocytes

2017 ◽  
Vol 53 (2) ◽  
pp. 167-173 ◽  
Author(s):  
May H. Hasan ◽  
Kamal G. Botros ◽  
Mona A. El-Shahat ◽  
Hussein A. Abdallah ◽  
Mohamed A. Sobh
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
In Vitro Differentiation
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