“…These pellets were consistently homogeneous and devoid of a “necrotic” or poorly cellularized center. The presence of cells and matrix in the pellet center is of particular interest because it provides evidence of a tolerance to hypoxia, if that is the milieu in the center of the pellet . This characteristic is considered an important feature of chondrocytes in normal articular cartilage where cells exist in a physiologic hypoxia…”
Section: Discussionmentioning
confidence: 99%
“…Whether this is a random pattern or is influenced by specific variables such as hydrostatic forces, oxygen tension, and another variable was not determined. It has been suggested that tissue culture in ambient oxygen concentrations may be suboptimal for chondrogenic induction …”
Objective: To determine the chondrogenic potential of cells derived from interzone tissue, the normal progenitor of articular cartilage during fetal development, compared to that of adult bone marrow-derived and adipose-derived mesenchymal cell isolates. The objective of this study was to compare the chondrogenic potential of fetal musculoskeletal progenitor cells to adult cell types, which are currently used therapeutically to facilitate joint cartilage repair in equine clinical practice. The hypothesis tested was that cells derived from interzone tissue have a chondrogenic potential that exceeds that of adult bone marrow-derived and adipose-derived mesenchymal cell isolates. Study design: In vitro study.Animals: Six young adult horses (15-17 months of age) and 6 equine fetuses aged 45-46 days of gestation. Methods: Three-dimensional pellet cultures were established under chondrogenic conditions with fresh, primary cells isolated from adult (articular cartilage, bone marrow, adipose, dermis) and fetal (interzone, skeletal anlagen cartilage, dermis) tissues.Cellular morphology, pellet architecture, and proteoglycan synthesis were assessed in the pellet cultures. Steady state levels of ACAN (aggrecan core protein), COL2A1(collagen type II), and COL1A1 (collagen type I) messenger RNA (mRNA) were compared among these cell types as pellet cultures and monolayer cultures. Results: Adult articular chondrocytes, fetal interzone cells, and fetal anlage cells generated the largest pellets under these chondrogenic culture conditions. Pellets derived from adult articular chondrocytes and fetal anlage cells had the highest scores on a neocartilage grading scale. Fetal anlage and adult articular chondrocyte pellets had low steady-state levels of COL1A mRNA but high COL2A1 expression. Anlage chondrocyte pellets also had the highest expression of ACAN. Conclusion: Adult articular chondrocytes, fetal interzone cells, and fetal anlage chondrocytes exhibited the highest chondrogenic potential. In this study, adult adipose-derived cells exhibited very limited chondrogenesis, and bone marrowderived cells had limited and variable chondrogenic potential.Clinical significance: Additional investigation of the high chondrogenic potential of fetal interzone cells and anlage chondrocytes to advance cell-based therapies in diarthrodial joints is warranted.
“…These pellets were consistently homogeneous and devoid of a “necrotic” or poorly cellularized center. The presence of cells and matrix in the pellet center is of particular interest because it provides evidence of a tolerance to hypoxia, if that is the milieu in the center of the pellet . This characteristic is considered an important feature of chondrocytes in normal articular cartilage where cells exist in a physiologic hypoxia…”
Section: Discussionmentioning
confidence: 99%
“…Whether this is a random pattern or is influenced by specific variables such as hydrostatic forces, oxygen tension, and another variable was not determined. It has been suggested that tissue culture in ambient oxygen concentrations may be suboptimal for chondrogenic induction …”
Objective: To determine the chondrogenic potential of cells derived from interzone tissue, the normal progenitor of articular cartilage during fetal development, compared to that of adult bone marrow-derived and adipose-derived mesenchymal cell isolates. The objective of this study was to compare the chondrogenic potential of fetal musculoskeletal progenitor cells to adult cell types, which are currently used therapeutically to facilitate joint cartilage repair in equine clinical practice. The hypothesis tested was that cells derived from interzone tissue have a chondrogenic potential that exceeds that of adult bone marrow-derived and adipose-derived mesenchymal cell isolates. Study design: In vitro study.Animals: Six young adult horses (15-17 months of age) and 6 equine fetuses aged 45-46 days of gestation. Methods: Three-dimensional pellet cultures were established under chondrogenic conditions with fresh, primary cells isolated from adult (articular cartilage, bone marrow, adipose, dermis) and fetal (interzone, skeletal anlagen cartilage, dermis) tissues.Cellular morphology, pellet architecture, and proteoglycan synthesis were assessed in the pellet cultures. Steady state levels of ACAN (aggrecan core protein), COL2A1(collagen type II), and COL1A1 (collagen type I) messenger RNA (mRNA) were compared among these cell types as pellet cultures and monolayer cultures. Results: Adult articular chondrocytes, fetal interzone cells, and fetal anlage cells generated the largest pellets under these chondrogenic culture conditions. Pellets derived from adult articular chondrocytes and fetal anlage cells had the highest scores on a neocartilage grading scale. Fetal anlage and adult articular chondrocyte pellets had low steady-state levels of COL1A mRNA but high COL2A1 expression. Anlage chondrocyte pellets also had the highest expression of ACAN. Conclusion: Adult articular chondrocytes, fetal interzone cells, and fetal anlage chondrocytes exhibited the highest chondrogenic potential. In this study, adult adipose-derived cells exhibited very limited chondrogenesis, and bone marrowderived cells had limited and variable chondrogenic potential.Clinical significance: Additional investigation of the high chondrogenic potential of fetal interzone cells and anlage chondrocytes to advance cell-based therapies in diarthrodial joints is warranted.
“…Along this line, preconditioning with hypoxia was shown to play a protective role in human stem cells, support their functions, and improve their therapeutic effect in vivo] [95][96][97]. Shell et al [98] demonstrated that equine ASCs cultured in the presence of 3% O 2 showed reduced proliferation compared to normoxic control conditions (21% O 2 ) . Furthermore, whereas hypoxia improved adipogenesis and chondrogenesis, the condition reduced osteogenic differentiation capacity [98].…”
Adipose-derived stem cells (ASCs) isolated from domestic animals fulfill the qualitative criteria of mesenchymal stem cells, including the capacity to differentiate along multiple lineage pathways and to self-renew, as well as immunomodulatory capacities. Recent findings on human diseases derived from studying large animal models, have provided evidence that administration of autologous or allogenic ASCs can improve the process of healing. In a narrow group of large animals used in bioresearch studies, pigs and horses have been shown to be the best suited models for study of the wound healing process, cardiovascular and musculoskeletal disorders. To this end, current literature demonstrates that ASC-based therapies bring considerable benefits to animal health in both spontaneously occurring and experimentally induced clinical cases. The purpose of this review is to provide an overview of the diversity, isolation, and characterization of ASCs from livestock. Particular attention has been paid to the functional characteristics of the cells that facilitate their therapeutic application in large animal models of human disease. In this regard, we describe outcomes of ASCs utilization in translational research with pig and horse models of disease. Furthermore, we evaluate the current status of ASC-based therapy in veterinary practice, particularly in the rapidly developing field of equine regenerative medicine. In conclusion, this review presents arguments that support the relevance of animal ASCs in the field of regenerative medicine and it provides insights into the future perspectives of ASC utilization in animal husbandry.
Graphical abstract
“…The cells were incubated at 37 °C, 5 % CO 2 and 90 % humidity up to con uency. In the current study, MSCs were characterized either by ow cytometry using the surface marker CD90 also using PCR for the expression of stem cell markers CD90, CD 105 and Oct 4 as previously reported by our group [28,29]. Upon con uency, cells were detached using TryplE (Gibco ® life technologies), were counted using a hemocytometer and were stored in 1 mL aliquots at -160 °C.…”
Section: Isolation Of Adipose Tissue Derived Mscsmentioning
AbstractBackground: Combination of mesenchymal stem cells (MSCs) and biomaterials is a rapidly growing approach in regenerative medicine particularly for chronic degenerative disorders including osteoarthritis and osteoporosis. In the present study, the effect of biomaterial bone substitutes on equine adipose derived MSCs morphology, viability, adherence, migration and osteogenic differentiation were investigated. Methods: MSCs were cultivated in conjunction with collagen CultiSpher-S Microcarrier (MC), nanocomposite xerogels B30 and B30Str biomaterials in osteogenic differentiate medium either under static or mechanical fluid shear stress (FSS) culture conditions. The data were generated by histological means, life cell imaging, cell viability, adherence and migration assays. Osteogenic differentiation was detected by semi-quantification of alkaline phosphatase (ALP) activity, matrix mineralization using Alizarin Red S (ARS) staining and quantification of the osteogenic markers; runt related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) expression using RT-qPCR. All data were statistically analyzed using ANOVA. Results: The data revealed that combined mechanical stress with MC but not B30 enhanced MSCs viability and promoted their migration. Combined osteogenic medium with MC, B30 and B30Str increased ALP activity compared to cultivation in basal medium. Osteogenic induction with MC, B30 and B30Str resulted in diffused matrix mineralization by means of ARS. FSS increased the viability in the presence of the osteogenic medium with MC but not B30 or B30Str. FSS enhanced osteogenic differentiation in the presence of B30Str. Upregulation of Runx2 and ALP expression was detected with osteogenic differentiation together with B30 and B30Str regardless of static or FSS culture. Conclusions: Taken together, the data revealed that FSS in conjunction with biomaterials promoted osteogenic differentiation of MSCs. This combination may be considered as a marked improvement for clinical applications to cure bone defects.
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