Immunophenotypic characterisation and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during <i>in vitro</i> culture

Mazurkevych, A. Y.; Malyuk, M. O.; Bezdieniezhnykh, Natalia; Starodub, L. F.; Kharkevych, Yuriy; Brusko, E. P.; Gryzińska, Magdalena; Jakubczak, Andrzej

doi:10.1515/jvetres-2016-0051

Cited by 7 publications

(7 citation statements)

References 25 publications

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“…However, BM-MSCs did not express N-Cadherin, another mesenchymal marker [18]. It can be speculated that BM-MSCs loose N-Cadherin expression during culture, since in another study regarding immunophenotypic characterization of equine BM-MSCs, it was observed that N-cadherin-positive cells were present at the second passage, whereas at the fifth passage these cells were not detected [19].…”

Section: Discussionmentioning

confidence: 99%

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

et al. 2019

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Background Little is known about the differences among adult and foetal equine mesenchymal stem cells (MSCs), and no data exist about their comparative ultrastructural morphology. The aim of this study was to describe and compare characteristics, immune properties, and ultrastructural morphology of equine adult (bone marrow: BM, and adipose tissue: AT) and foetal adnexa derived (umbilical cord blood: UCB, and Wharton’s jelly: WJ) MSCs. Results No differences were observed in proliferation during the first 3 passages. While migration ability was similar among cells, foetal MSCs showed a higher adhesion ability, forming smaller spheroids after hanging drop culture ( P < 0.05). All MSCs differentiated toward adipogenic, chondrogenic and osteogenic lineages, only tenogenic differentiation was less evident for WJ-MSCs. Data obtained by PCR confirmed MHC1 expression and lack of MHC2 expression in all four cell types. Foetal adnexa MSCs were positive for genes specific for anti-inflammatory and angiogenic factors (IL6, IL8, ILβ1) and WJ-MSCs were the only positive for OCT4 pluripotency gene. At immunofluorescence all cells expressed typical mesenchymal markers (α-SMA, N-cadherin), except for BM-MSCs, which did not express N-cadherin. By transmission electron microscopy, it was observed that WJ-MSCs had a higher ( P < 0.05) number of microvesicles compared to adult MSCs, and UCB-MSCs showed more microvesicles than BM-MSCs ( P < 0.05). AT-MSCs had a lower number of mitochondria than WJ-MSCs ( P < 0.05), and mitochondrial area was higher for WJ-MSCs compared to UCB and AT-MSCs ( P < 0.05). Conclusions Results demonstrate that MSCs from adult and foetal tissues have different characteristics, and foetal MSCs, particularly WJ derived ones, seem to have some charactestics that warrant further investigation into potential advantages for clinical application.

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Section: Discussionmentioning

confidence: 99%

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

et al. 2019

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“…In this context, a comparable or even higher genetic stability was already shown with hPL compared to FBS ( 23 , 24 ). However, the genetic stability of equine MSC has so far only been studied with FBS ( 22 , 26 – 28 ). For this reason, in the present study, we analyzed the genetic stability of equine AD-MSC cultivated with either FBS or ePL.…”

Section: Introductionmentioning

confidence: 99%

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

Hagen

Niebert²,

Brandt³

et al. 2022

Front. Vet. Sci.

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Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from n = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium (p < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells (p < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL (p < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties.

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“…Слід зазначити, що для лікування коня ми використовували стовбурові клітини третього пасажу, оскільки наші попередні дослідження показують, що рівень мінливості каріотипу мезенхімальних стовбурових клітин коней, зокрема, отриманих з кісткового мозку, на 3 пасажі культивування in vitro відповідає спонтанним рівням, характерним для цього виду тварин. Це свідчить про те, що ці клітини безпечні для терапевтичних цілей (Mazurkevych et al, 2016).…”

Section: матеріали і методиunclassified

The effect of mesenchymal stem cells and platelet-rich plasma on the course of chronic laminitis in horse: a clinical case

Malyuk¹,

Kharkevych²,

Brusko³

et al. 2021

VVTP

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Laminitis or rheumatic inflammation of the hooves is inflammation of the hoof lamina, in which the connection between the coffin bone and the horny shoe is disrupted as a result of the destruction of the connection between the leaves of the horny wall with the leaves of the base of the hoof skin. The pathogenesis of this disease is quite complex and contradictory. Some authors associate it with the accumulation of biogenic amines in the hoof tissues, while others point to the violation of lamellar microcirculation and the effect of tissue metalloproteinases on the leaves of the horny wall and the base of the hoof skin. Treating horses with laminitis is not an easy task, due to the complex pathogenesis of the disease. Thus, the search for modern treatments of horses with laminitis is an urgent and timely task. The aim of our study was to investigate the effect of adipose-derived mesenchymal stem cells and platelet-rich plasma on the course of chronic laminitis in horses. The object of the study was a castrated stallion of a Ukrainian riding breed, 8 years old, with a clinical manifestation of chronic laminitis with detachment of the hoof wall along the border line on the left forelimb. Materials and methods. Autologous mesenchymal stem cells were obtained from adipose tissue. A culture of mesenchymal stem cells of adipose tissue in the amount of 12 million cells was injected into the palmar medial digital artery of the injured limb. The introduction was repeated three times, while the cells were suspended in 3 cm3 of phosphate-buffered saline. The third injection of cells was performed in combination with autologous platelet-rich plasma. Results: Active recovery of the hoof wall was observed on the injured forelimb. Coronary fistulas and damage of the hoof joint capsule were healed. The shape of the hoof wall and sole began to normalize. The pulsation of the arteries of the toes has become less pronounced. No corona edema was observed in the area. Conclusion. Under the influence of transplanted mesenchymal stem cells and platelet-rich blood plasma, tissue structures of the injured hoof had restored as the clinical and functional state of the injured limb; at the same time, the x-ray picture of the hoof worsens, which is accompanied by irreversible processes in the hoof tissues and its deformation.

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Immunophenotypic characterisation and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture

Cited by 7 publications

References 25 publications

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

The effect of mesenchymal stem cells and platelet-rich plasma on the course of chronic laminitis in horse: a clinical case

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