Isolation and characterization of mesenchymal stem cells derived from bovine Wharton's jelly and their potential for use in cloning by nuclear transfer

Silva, Carolina Gonzales da; Martins, Claudia Miranda; Cardoso, Tereza Cristina; Cunha, Elisa Ribeiro da; Bessler, H. C.; McManus, Concepta; Pivato, I.; Báo, Sônia Nair

doi:10.1590/0103-8478cr20151074

Cited by 12 publications

(10 citation statements)

References 19 publications

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“…UCs used for different experiments are commonly collected at the time of parturition [ 14 , 16 ]. Here, in this study, the UCs were obtained from the nearby veterinary hospitals with informed consent of the owners and by following all the safety compliances of the IAEC.…”

Section: Methodsmentioning

confidence: 99%

“…The in vitro explant tissue culture is a reliable method for easy isolation of cells [ 14 , 16 ]. The UCs were carefully washed in sterile chilled phosphate-buffered saline (PBS) to rinse off the blood and blood clots within it, if any.…”

Section: Methodsmentioning

confidence: 99%

“…The explant culture of UCs for the isolation of MSCs has been performed for different in vitro cellular experiments [ 14 , 16 ]. Herein, the explant culture was maintained in vitro for around 10 days.…”

Section: Methodsmentioning

confidence: 99%

“…[ 15 ] and Silva et al . [ 16 ] showed the possibility of isolating these cells by in vitro culture system. However, to make the cells more accessible on large scale by minimizing ethical concern, it is necessary to standardize a non-invasive isolation technique of bMSCs.…”

Section: Introductionmentioning

confidence: 99%

“…Cardoso et al [14] made an attempt to isolate bMSCs from Wharton's jelly (WJ) and maintained in 3D serumfree condition which guided the scientific community toward utilizing cord tissue as a readily accessible potential source of bMSCs. Later on, Xiong et al [15] and Silva et al [16] showed the possibility of isolating these cells by in vitro culture system. However, to make the cells more accessible on large scale by minimizing ethical concern, it is necessary to standardize a non-invasive isolation technique of bMSCs.…”

Section: Introductionmentioning

confidence: 99%

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Post-calving umbilical cord tissue offcut: A potential source for the isolation of bovine mesenchymal stem cells

et al. 2020

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Background and Aim: Veterinary health care is an emergent area in animal sciences and innovative therapeutic approaches happen to be imperative in the present days. In view of the importance of cattle health and production, it is necessary to take up contemporary approach of stem cell therapy in this sector also. This study aimed to standardize an explant culture method of bovine umbilical tissue offcut to isolate mesenchymal stem cells (MSCs) because considerable efforts are required for ensuring easy accessibility and availability of MSCs in bulk quantity, as well as in establishing and characterizing the cell lines. Materials and Methods: The umbilical cord (UC) tissue matrix offcut was collected after calving. A simplified in vitro cell isolation technique was followed to collect the emerged out cells from the explants of UC. Further, we expanded these isolated cells in vitro, observed its growth kinetics, and characterized to confirm as per the criterion of bovine MSCs. Results: A considerable exponential growth rate of the UC-derived cells was noticed. In addition to their confirmation as MSCs, the cells also exhibited plastic adherent property and maintained the spindle-shaped morphology throughout the in vitro culture. The cultured cells were found positive MSC-specific surface markers CD105, CD90, and CD73 and were negative for hematopoietic cell marker CD45. Cytochemical studies revealed the ability of the cells to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Conclusion: This simplified method of isolation and culture of bovine multipotent MSCs from the UC offcut collected after calving could be extrapolated for the greater availability of the cells for prospective therapeutic applications.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Post-calving umbilical cord tissue offcut: A potential source for the isolation of bovine mesenchymal stem cells

et al. 2020

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Validation of multiparametric panels for bovine mesenchymal stromal cell phenotyping

et al. 2023

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Bovine mesenchymal stromal cells (MSCs) display important features that render them valuable for cell therapy and tissue engineering strategies, such as self‐renewal, multi‐lineage differentiation, as well as immunomodulatory properties. These cells are also promising candidates to produce cultured meat. For all these applications, it is imperative to unequivocally identify this cell population. The isolation and in vitro tri‐lineage differentiation of bovine MSCs is already described, but data on their immunophenotypic characterization is not yet complete. The currently limited availability of monoclonal antibodies (mAbs) specific for bovine MSC markers strongly hampers this research. Following the minimal criteria defined for human MSCs, bovine MSCs should express CD73, CD90, and CD105 and lack expression of CD14 or CD11b, CD34, CD45, CD79α, or CD19, and MHC‐II. Additional surface proteins which have been reported to be expressed include CD29, CD44, and CD106. In this study, we aimed to immunophenotype bovine adipose tissue (AT)‐derived MSCs using multi‐color flow cytometry. To this end, 13 commercial Abs were screened for recognizing bovine epitopes using the appropriate positive controls. Using flow cytometry and immunofluorescence microscopy, cross‐reactivity was confirmed for CD34, CD73, CD79α, and CD90. Unfortunately, none of the evaluated CD105 and CD106 Abs cross‐reacted with bovine cells. Subsequently, AT‐derived bovine MSCs were characterized using multi‐color flow cytometry based on their expression of nine markers. Bovine MSCs clearly expressed CD29 and CD44, and lacked expression of CD14, CD45, CD73, CD79α, and MHCII, while a variable expression was observed for CD34 and CD90. In addition, the mRNA transcription level of different markers was analyzed using reverse transcription quantitative polymerase chain reaction. Using these panels, bovine MSCs can be properly immunophenotyped which allows a better characterization of this heterogenous cell population.

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Mesenchymal stem cell: Basic research and potential applications in cattle and buffalo

Gugjoo

Amarpal

Fazili

et al. 2018

Journal Cellular Physiology

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Characteristic features like self‐renewal, multilineage differentiation potential, and immune‐modulatory/anti‐inflammatory properties, besides the ability to mobilize and home distant tissues make stem cells (SCs) a lifeline for an individual. Stem cells (SCs) if could be harvested and expanded without any abnormal change may be utilized as an all‐in‐one solution to numerous clinical ailments. However, slender understanding of their basic physiological properties, including expression potential, behavioral alternations during culture, and the effect of niche/microenvironment has currently restricted the clinical application of SCs. Among various types of SCs, mesenchymal stem cells (MSCs) are extensively studied due to their easy availability, straightforward harvesting, and culturing procedures, besides, their less likelihood to produce teratogens. Large ruminant MSCs have been harvested from various adult tissues and fetal membranes and are well characterized under in vitro conditions but unlike human or other domestic animals in vivo studies on cattle/buffalo MSCs have mostly been aimed at improving the animals’ production potential. In this document, we focused on the status and potential application of MSCs in cattle and buffalo.

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Isolation and characterization of mesenchymal stem cells derived from bovine Wharton's jelly and their potential for use in cloning by nuclear transfer

Cited by 12 publications

References 19 publications

Post-calving umbilical cord tissue offcut: A potential source for the isolation of bovine mesenchymal stem cells

Post-calving umbilical cord tissue offcut: A potential source for the isolation of bovine mesenchymal stem cells

Validation of multiparametric panels for bovine mesenchymal stromal cell phenotyping

Mesenchymal stem cell: Basic research and potential applications in cattle and buffalo

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