Isolation and expansion of equine umbilical cord‐derived matrix cells (EUCMCs)

Passeri, Simona; Nocchi, Francesca Romana; Lamanna, Roberta; Lapi, Simone; Miragliotta, Vincenzo; Giannessi, Elisabetta; Abramo, Francesca; Stornelli, Maria Rita; Matarazzo, Micheletino; Plenteda, Daniele; Urciuoli, Patrizia; Scatena, F.; Coli, Alessandra

doi:10.1016/j.cellbi.2008.10.012

Cited by 48 publications

(35 citation statements)

References 33 publications

(45 reference statements)

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“…Further investigation using RT-PCR is needed to verify if different differentiation capacities exist among these three cell lines. (Passeri et al 2009, Lovati et al 2011. The isolation rate obtained in the present study by centrifuging UCB on a Percoll densitygradient was lower than that obtained by Koch et al (2009) using a PrepaCyte-EQ density gradient (100%).…”

Section: In Vitro Differentiationcontrasting

confidence: 40%

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

Iacono

Brunori²,

Pirrone³

et al. 2012

Reproduction

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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 (2.0G0.6 days) than those isolated from CB (2.6G1.3 days) and AF (2.3G1.0 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.

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Section: In Vitro Differentiationcontrasting

confidence: 40%

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

Iacono

Brunori²,

Pirrone³

et al. 2012

Reproduction

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“…More recently, primitive MSCs have also been isolated from equine UCM [9][10][11][12]. Specific pluripotent and MSCmarkers of equine and swine UCM are reported in Table 1.…”

Section: Introductionmentioning

confidence: 99%

“…Due to their primitive embryonic stem-cell-like characteristics, UCM-derived cells, when exposed to specific stimuli, have the ability to differentiate into multiple germ layers including mesoderm in horses [9,10,13] and ectoderm in pigs [8,14] and horses [9,11]. Others: SOX2+ [9][10][11][12] Swine Embryonic cell markers: OCT-3/4+ Others: SOX2+, NANOG+ [8] As in human medicine (for review see [7,15,16]), also in veterinary medicine an important property to evaluate stem cells is to assess their usefulness in allogenic regenerative medicine. To test the immunological properties of UCM-derived cells, Carrade et al, (2010) studied healthy horses following a single intra-articular injection of autologous and allogeneic MSCs from equine UCM and found no significant differences between the degree and type of inflammation elicited by self and non-self-MSC [17].…”

Section: Introductionmentioning

confidence: 99%

In Vitro Studies of Horse Umbilical Cord Matrix-Derived Cells: From Characterization to Labeling for Magnetic Resonance Imaging

Consiglio

Corradetti

Rutigliano

et al. 2011

TOTERMJ

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Despite the increasing use of cell-based therapies for equine orthopedic problems, many questions remain to be answered, such as what is: the optimal cell source for particular injuries, the best timing and route of treatment and the long-term safety and efficacy of the procedure? Previously, equine mesenchymal stem cells (MSCs) have most frequently been isolated from bone marrow (BM) and adipose tissue. However, these cells have limited potential in terms of in vitro proliferation ability and differentiation capacity with increasing donor age and in vitro passage number. In addition, procurement of BM-derived cells in horses requires an invasive BM aspiration procedure which has been associated with pneumopericardium. Fetal adnexa could provide a useful alternative source of MSCs avoiding these limitations. To investigate this, we isolated and characterized presumptive stem cells from the intervascular and perivascular portions of equine umbilical cord matrix using enzymatic digestion. The cells isolated from the intervascular portion showed faster doubling times than cells from the perivascular portion (which are probably more highly differentiated). Cells from both portions expressed MSC mRNA markers (CD29, CD105, CD44, CD166) and were negative for CD34 and MHC-II. Osteogenic, adipogenic, chondrogenic and neurogenic differentiation were confirmed by specific staining and gene expression.To investigate the potential of umbilical cord-derived cells for use in cell therapies, pre-clinical experiments involving labeling of cells with magnetic resonance contrast agents (superparamagnetic iron oxide particles -SPIO -and manganese chloride) and the subsequent in vitro study of these were conducted. The SPIO labeling procedure proved to be an efficient and non toxic tool that merits further investigation and the possible development of in vivo studies.

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“…As such, gestational tissues including umbilical cord blood (Kern et al 2006, Koch et al 2007, Guest 2008, Reed & Johnson 2008, Bartholomew et al 2009, Schuh et al 2009, Seo et al 2009, Raoufi et al 2011, umbilical cord tissues (Mitchell et al 2003, Carlin et al 2006, Hoynowski et al 2007, Cremonesi et al 2008, Passeri et al 2009, Lovati et al 2011, Iacono et al 2012a, amniotic tissues (Filioli Uranio et al 2011, Iacono et al 2012a, 2012b, Lange-Consiglio et al 2012, and amniotic fluid (AF) (Chen et al 2011, Dev et al 2011, Lovati et al 2011, Iacono et al 2012a) have been recently suggested, also in veterinary medicine, as appealing candidates for the derivation of MSCs to use in cell biotechnology. The human lesson teaches that presumptive adult stem cells derived from gestational tissues retain highest proliferation capacity, longest telomere length, broadest differentiation, and extensive proliferative potential when compared with cells obtained from adult tissues (Kogler et al 2004, Kern et al 2006.…”

Section: Introductionmentioning

confidence: 99%

Mesenchymal stem cells from amnion and amniotic fluid in the bovine

Corradetti¹,

Meucci²,

Bizzaro³

et al. 2013

Reproduction

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Amnion and amniotic fluid (AF) are noncontroversial and inexhaustible sources of mesenchymal stem cells (MSCs) that can be harvested noninvasively at low cost. As in humans, also in veterinary field, presumptive stem cells derived from these tissues reveal as promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. The aim of this work is to obtain and characterize, for the first time in bovine species, presumptive MSCs from the epithelial portion of the amnion (AECs) and from the AF (AF-MSCs) to be used for clinical applications. AECs display a polygonal morphology, whereas AF-MSCs exhibit a fibroblastic-like morphology only starting from the second passage, being heterogeneous during the primary culture. For both lines, the proliferative ability has been found constant over the ten passages studied and AECs show a statistically lower (P!0.05) doubling time with respect to AF-MSCs. AECs express MSC-specific markers (ITGB1 (CD29), CD44, ALCAM (CD166), ENG (CD105), and NT5E (CD73)) from P1 to P3; in AF-MSCs, only ITGB1, CD44, and ALCAM mRNAs are detected; NT5E is expressed from P2 and ENG has not been found at any passage. AF-MSCs and AECs are positive for the pluripotent markers (POU5F1 (OCT4) and MYC (c-Myc)) and lack of the hematopoietic markers. When appropriately induced, both cell lines are capable of differentiating into ectodermal and mesodermal lineages. This study contributes to reinforce the emerging importance of these cells as ideal tools in veterinary medicine. A deeper evaluation of the immunological properties needs to be performed in order to better understand their role in cellular therapy.

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Isolation and expansion of equine umbilical cord‐derived matrix cells (EUCMCs)

Cited by 48 publications

References 33 publications

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

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

In Vitro Studies of Horse Umbilical Cord Matrix-Derived Cells: From Characterization to Labeling for Magnetic Resonance Imaging

Mesenchymal stem cells from amnion and amniotic fluid in the bovine

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