Isolation and multilineage differentiation of bovine bone marrow mesenchymal stem cells

Abstract

The bone marrow harbors a population of mesenchymal stem cells (MSCs) that possess the potential to differentiate into bone, cartilage, and fat, and along other tissue pathways. To date, MSCs from various species have been studied. Despite the bovine experimental model being widely used in experiments in vivo and in vitro, only a limited amount of information regarding bovine MSCs is available. The aim of this study was to isolate and induce the multilineage mesenchymal differentiation of bovine MSCs, thereby initiating further research on these cells. Bovine MSCs were isolated from eight calves, and osteogenic, chondrogenic, and adipogenic differentiation was induced by using a combination of previously reported protocols for other species. The level of differentiation was evaluated by histological examination and by analyzing the expression of tissue-specific genes by a quantitative "real time" reverse transcription/polymerase chain reaction technique. Following osteoinduction, the isolated fibroblast-like cells transformed into cuboidal cells and formed alkaline-phosphatase-positive colonies; during differentiation, these colonies transformed into mineralized nodules. In addition, osteogenesis was followed by osteocalcin and collagen type I mRNA expression. Chondrogenesis was confirmed by the demonstration of collagen type II, aggrecan, and sox9 mRNA expression in the cells stimulated by transforming growth factor β1 in monolayer culture. After being cultured in an adipogenesis-inducing medium, the MSCs responded by the accumulation of lipid vacuoles and the expression of adipocyte-specific genes. We have therefore demonstrated that cells harvested from bovine bone marrow are capable of in vitro extensive multiplication and multilineage differentiation, making them a relevant and invaluable model in the field of stem cell research.

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

“…The results from our previous (Bosnakovski et al, 2005) and present studies clearly indicate that MSCs cultured under various culture conditions and treated with TGF b1 undergo chondrogenic differentiation, which is time-and dose-dependent with respect to the stimulants. TGF b1 exerts its effect through a high-affinity interaction with a heteromeric receptor complex (type I and II) that comprises two structurally related serinethreonine kinases.…”

“…Using a bovine experimental model, we have previously analyzed MSC chondrogenesis in monolayer cultures in response to TGF b1 stimulation (Bosnakovski et al, 2005). We also demonstrated that bovine MSC has the potential for self-inducing chondrogenic differentiation independent of any externally added biostimulants, if strong cell-cell interaction in the condensate pellet culture system is allowed (Bosnakovski et al, 2004).…”

“…Bone marrow was aspirated from four calves (2 months old), and MSCs were isolated by previously described methods (Bosnakovski et al, 2004(Bosnakovski et al, , 2005. Briefly, the bone marrow sample was washed twice with phosphate buffered saline (PBS), and twice again with Dulbecco's Modified Eagle Medium (DMEM; GIBCO BRL, Grand Islands, NY).…”

Bone Marrow-Derived Mesenchymal Stem cells Infusion Ameliorates Hyperglycemia, Dyslipidemia, Liver and Kidney Functions in Diabetic Rats

“…In agreement of our study, the result of Abdel Aziz et al [72] (2014) showed a significant improvement in kidney function (serum urea and creatinine) in the diabetic nephropathy (DN) group treated intravenously with single dose of 10 6 MSCs per rat compared to DN group. They referred this improvement in kidney function in MSCs treated group to their paracrine action via different growth factors such as VEGF, TGF-β and TNF-α and antiapoptotic effects via Bax and Bcl2 genes.…”

“…The bone marrow in addition to haemopoietic stem cells comprises a population of mesenchymal stem cells, these cells exhibit multilineage differentiation capacity [6]. By using MSCs, we are thus using the "mother cell" from which pancreas and other tissues were originally derived [7], [8].…”