Ovine bone marrow mesenchymal stem cells: isolation and characterization of the cells and their osteogenic differentiation potential on embroidered and surface-modified polycaprolactone-co-lactide scaffolds

Abstract

The current study was undertaken with the goal being isolation, cultivation, and characterization of ovine mesenchymal stem cells (oMSC). Furthermore, the objective was to determine whether biological active polycaprolactone-co-lactide (trade name PCL) scaffolds support the growth and differentiation of oMSC in vitro. The oMSC were isolated from the iliac crest of six merino sheep. Three factors were used to demonstrate the MSC properties of the isolated cells in detail. (1) Their ability to proliferate in culture with a spindle-shaped morphology, (2) presence of specific surface marker proteins, and (3) their capacity to differentiate into the three classical mesenchymal pathways, osteoblastic, adipogenic, and chondrogenic lineages. Furthermore, embroidered PCL scaffolds were coated with collagen I (coll I) and chondroitin sulfate (CS). The porous structure of the scaffolds and the coating with coll I/CS allowed the oMSC to adhere, proliferate, and to migrate into the scaffolds. The coll I/CS coating on the PCL scaffolds induced osteogenic differentiation of hMSC, without differentiation supplements, indicating that the scaffold also has an osteoinductive character. In conclusion, the isolated cells from the ovine bone marrow have similar morphologic, immunophenotypic, and functional characteristics as their human counterparts. These cells were also found to differentiate into multiple mesenchymal cell types. This study demonstrates that embroidered PCL scaffolds can act as a temporary matrix for cell migration, proliferation, and differentiation of oMSC. The data presented will provide a reliable model system to assess the translation of MSC-based therapy into a variety of valuable ovine experimental models under autologous settings.

Isolation and Characterization of CD271⁺ Stem Cells Derived from Sheep Dermal Skin

“…Ovine bmMSCs have been reported to differentiate into adipocytes [Perry et al, 2003] and osteocytes [Rentsch et al, 2010]. We could confirm these results also for CD271-dMSCs.…”

Hard Tissue Augmentation of Aged Bone by Means of a Tin-Free PLLA-PCL Co-Polymer Exhibiting in vivo Anergy and Long-Term Structural Stability

“…Prior to applying the explanted cells in animal experiments the primary ovine bone marrow-derived MSC were evaluated in culture. The cells revealed a spindle-like morphology with long processes and showed indistinguishable cell proliferation and colony formation when cultivated at 3% or 20% O 2 , yet exhibited slower rates when compared to human bone marrow-derived MSC [15, 16]. Colony formation was determined by seeding cells in 58 cm 2 cell culture dishes at a density of 10 cells/cm 2 .…”

“…They were positive for CD44 and found negative for CD45, CD11b, CD31, CD14, and CD105, and exhibited tri-lineage differentiation potential. Assays revealing cellular differentiation were performed according to Rentsch et al [16]. Osteogenic differentiation was induced by adding 0.5 µM L-ascorbic acid phosphate (Fluka #49752), 1 mM β-glycerol phosphate (Fluka #50020), and 100 nM dexamethasone (Sigma #D4902) to standard growth medium.…”

Therapeutic potential of stem cells in veterinary practice

“…Further, Caplan (1991) for Vetstem (U.S.A., http://www.vet-stem.com/), Medistem the first time named these cells as mesenchymal stem Inc. (U.S.A., http://medisteminc.com/), Histostem cells. MSCs have been isolated from a number of (South Korea, http://www.histostem.co.kr) provide species like human, rat, mice, dog, cat, pig, horse, stem cell therapy for animals and claims cure for sheep, goat, cattle [11,[38][39][40][41][42][43][44][45][46]. MSCs can be isolated orthopedic and other injuries (for websites see reference).…”