scholarly journals Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Reproduction ◽  
2012 ◽  
Vol 143 (4) ◽  
pp. 455-468 ◽  
Author(s):  
Eleonora Iacono ◽  
Lara Brunori ◽  
Alessandro Pirrone ◽  
Pasquale Paolo Pagliaro ◽  
Francesca Ricci ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Cell Lines ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Wharton’S Jelly ◽  
Blue Staining ◽  
Wharton's Jelly

Mesenchymal stem cells (MSCs) have been derived from multiple sources of the horse including umbilical cord blood (UCB) and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), CB and Wharton's Jelly (WJ). Samples were obtained from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was prepared by incubating with an enzymatic solution for 2 h. All cell lines were cultured in DMEM/TCM199 plus fetal bovine serum. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB and 8/12 (66.7%) WJ samples. Statistically significant differences were found between cell-doubling times (DTs): cells isolated from WJ expanded more rapidly (2.0±0.6 days) than those isolated from CB (2.6±1.3 days) and AF (2.3±1.0 days) (P<0.05). Positive von Kossa and Alizarin Red S staining confirmed osteogenesis. Alcian Blue staining of matrix glycosaminoglycans illustrated chondrogenesis and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105; and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from a uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their use as an allogenic cell source, further studies are needed for evaluating the expression of markers related to cell immunogenicity.

Wharton's jelly derived mesenchymal stem cells: regenerative medicine beyond umbilical cord blood

Placenta ◽  
2011 ◽  
Vol 32 ◽  
pp. S339 ◽  
Author(s):  
R.R. Taghizadeh ◽  
K.E. Pollok ◽  
M. Betancur ◽  
L. Boissel ◽  
K.J. Cetrulo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Wharton’S Jelly ◽  
Wharton's Jelly

scholarly journals Potency of umbilical cord blood- and Wharton’s jelly-derived mesenchymal stem cells for scarless wound healing

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Hanako Doi ◽  
Yuriko Kitajima ◽  
Lan Luo ◽  
Chan Yan ◽  
Seiko Tateishi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Wharton’S Jelly ◽  
Wharton's Jelly

384 ISOLATION OF MESENCHYMAL STEM CELLS FROM WHARTON'S JELLY, CORD BLOOD, AND AMNIOTIC FLUID IN THE HORSE

2010 ◽  
Vol 22 (1) ◽  
pp. 348 ◽  
Author(s):  
E. Iacono ◽  
L. Brunori ◽  
A. Pirrone ◽  
B. Merlo
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Culture Medium ◽  
Wharton’S Jelly ◽  
Population Doubling ◽  
Blue Staining ◽  
Wharton's Jelly ◽  
Orthopedic Injuries ◽  
Alternative Sources

The effect of orthopedic injuries on the health of racehorse and racing financials can be dramatic. Bearing this in mind, there is naturally a great interest in new possible treatments for orthopedic injuries. In this study we evaluated equine amniotic fluid (AF), cord blood (CB), and Wharton’s jelly (WJ) as alternative sources of primitive, multipotent stem cells. All chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated. Samples were recovered at labor from 7 standardbred mares with ages between 6 and 15 years. Wharton’s jelly was incubated with collagenasis at 37°C for at least 2 hr. The resulting cell suspension was centrifuged, and the cell pellet was resuspended in culture medium (DMEM and TCM-199, 1:1) plus 10% FBS (GIBCO®, Invitrogen Corporation, Carlsbad, CA, USA), 100 IU mL-1 penicillin, and 100 μg mL-1 streptomycin). Amniotic fluid and CB were diluted 1:1 in PBS medium supplemented with antibiotic solution and centrifuged for 15 min at 300 x g. The pellet was resuspended in 5 mL of culture medium, placed on 5 mL of 70% Percoll solution, and centrifuged at 1200 x g for 30 min. The interface layer was then aspirated and resuspended in culture medium. All the cell lineages were washed seeded into 25 cm2 flasks and cultured in a 5% CO2 incubator at 38.5°C. Medium was refreshed after 48 h and then twice a week. Adherent fibroblast-like cells were tested for their differentiation potential at passage 3. Chondrogenic differentiation was performed using a micromass culture technique, whereas osteogenic differentiation was induced in monolayer culture (Mizuno H and Hyakusoku H 2003 J. Nippon Med. Sch. 70, 300-306). Evidence for differentiation was made via morphological, biochemical, and cytological evaluations. Sample volume was 15-55 mL for AF, 10-60 mL for CB, and 5-12.6 g for WJ. In 4 out of 6 (66.7%) AF, 5 out of 5 (100%) CB, and 6 out of 7 (85.7%) WJ samples, colonies with the classical mesenchymal stem cell (MSC) morphology of adherent fibroblastoid spindle-shaped cells growing in a monolayer were observed. Undifferentiated cells have been passaged up to 8 times, population-doubling times (DT) were calculated for a subset of cell passages, and data were analyzed by ANOVA (Statistica for Windows, Stat Soft Inc., Tulsa, OK, USA). In all samples, there were no significant differences (P > 0.05) between DT of all passages. The DT was greater (P < 0.05) forCB (2.6 ± 1.3 d) than for AF (2.1 ± 0.8 d) and WJ (1.9 ± 0.5 d). By passage 8, the cells had undergone 39.1 ± 1.2 cell-doubling numbers in AF, 35.1 ± 2.7 in CB, and 37.4 ± 2.0 in WJ. A spheroid structure and positive Alcian blue staining of matrix glycosaminoglycans illustrated chondrogenesis. Cellular morphology and positive von Kossa staining, as well as alkaline phosphatase activity, confirmed osteogenesis. Based on the results obtained, WJ, CB, and for the first time in the horse, AF represent alternative sources of stem cells. Their differentiation into osteocytes and chondrocytes demonstrated that equine MSC from these samples can be induced to form multiple cell types, which underlies their value for regenerative medicine in injured horses.

407: Characterization of fetal mesenchymal stem cells isolated from umbilical cord blood and amniotic fluid of the same donor

2009 ◽  
Vol 201 (6) ◽  
pp. S157
Author(s):  
Francesca Maria Russo ◽  
Patrizia Vergani ◽  
Francesca Gatto ◽  
Simona Marzorati ◽  
Ilaria Follesa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood
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