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Mesenchymal stem cells are self-renewing capacity with multi- differentiation potential of progenitor cell. The purpose of this study was to isolate, proliferate and morphological characterization of mesenchymal stem cells (MSC) collected from bone marrow of different species of animals. Bone marrow aspiration was collected with the help of a bone...
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Background
In an attempt to increase the therapeutic potential for myocardial regeneration, there is a quest for new cell sources and types for cell therapy protocols. The pathophysiology of heart diseases may affect cellular characteristics and therapeutic results. Methods
To study the proliferative and differentiation potential of mesenchymal ste...
Citations
... Biopsy needle stylet was removed and 5 ml of BMA was collected in hypodermic syringe containing anticoagulant. Later this BMA was used at fractured site (Maiti et al., 2013). ...
... At 70-80% confluence, 0.25% trypsin-EDTA (Gibco, USA) was used to detach the cells that were sub-cultured at the ratio of 1:2. Experiments were carried out using third-passage cells, then isolated cells were identified by their plastic adherent capability and the shape of their spindles (elongated shape) which is detected by an inverted microscope [14] After 3 rd passage, the BM-MSCs were washed 2 times with PBS, resuspended in culture media, and intravenously infused after AMI confirmation [15]. ...
... Very broadly they comprise of two major classes: embryonic and adult mesenchymal stem cells. 1 Mesenchymal stem cells (MSCs) because of their self replication, differentiate into various types of mature cells and tissues, and regeneration capabilities are regarded as an excellent source of cells for tissue engineering and for treatment of various incurable diseases and therapeutic uses in gene therapy, drug delivery, and reconstructive surgery. 2,3 Recently, induced pluripotent stem cells (iPSC) and embryonic stem cells (ESCs) attracted researchers in organogenesis and cellmediated therapy experiments, however, teratoma formation, ethical issues, and graft vs host rejection are the major limitations in development and therapeutic application of these cells. ...
Equine multipotent mesenchymal stem cells (hrs-MSC) can be isolated from various tissues including adipose tissue (AT). We have analyzed the effect of different serum sources on hrs-AT-MSC cultured and assessed proliferation, morphology, viability and immunophenotype and plasticity. The hrs-AT-MSC was cultured separately with growth media containing three different serums: 20% FCS (Gibco), 20% horse serum (Sigma) and 20% allogenic horse serum (Belgium lab) in CO2 incubators. The hrs-AT-MSC growth and proliferation was better in cultural conditions where 20% FCS and 20% horse serum (Sigma) were used. Mesenchymal stem cell count was highest in the condition where horse serum (sigma) was used than both FCS and horse allogenic serum. The viability was more in where allogenic serum (Belgium lab) was used than both FCS (Gibco) and horse serum (Sigma). Like FCS (Gibco), horse serum (Sigma) and allogenic horse serum (Belgium lab) also showed promising /positive effects on equine adipose tissue derived mesenchymal stem cell (hrs-At-MSC) culture and proliferation. Horse serum was found as efficient as fetal calf serum in supporting proliferation and differentiation of equine mesenchymal stem cells in vitro. Further studies are needed to analyze these aspects of MSC in tissue regeneration. Stem cell biology has attracted tremendous interest recently. It is hoped that it will play a major role in the treatment of a number of incurable diseases via transplantation therapy. Several verities of stem cells have been isolated and identified in vivo and in vitro. Very broadly they comprise of two major classes: embryonic and adult mesenchymal stem cells.1 Mesenchymal stem cells (MSCs) because of their self replication, differentiate into various types of mature cells and tissues, and regeneration capabilities are regarded as an excellent source of cells for tissue engineering and for treatment of various incurable diseases and therapeutic uses in gene therapy, drug delivery, and reconstructive surgery.2,3 Recently, induced pluripotent stem cells (iPSC) and embryonic stem cells (ESCs) attracted researchers in organogenesis and cell-mediated therapy experiments, however, teratoma formation, ethical issues, and graft vs host rejection are the major limitations in development and therapeutic application of these cells.4 Due to these limitations, mesenchymal stem cells (MSCs) from adult tissues are now attractive material for and tissue engineering and cell-mediated therapy.5 Isolation of MSC derived from equine species has been reported in a number of different tissues, including bone marrow,6 peripheral blood,7 fat tissue8 and umbilical cord blood.9 Adipose tissue derived equine MSC (hrs-AT-MSC) exhibit the ability to differentiate into different types of cells and tissues in appropriate culture conditions using growth factors and specific hormones into osteoblast, chondroblat and adipocytes and a profound proliferative ability without hampering their own genetic firmness.8 Serum is an integral component for MSC culture in vitro and also it is essential for osteogenic differentiation of MSC along with other factors includes β glycerophosphate, ascorbic acid, and dexamethasone as osteogenic supplements. 10 Serum is an essential component of complete growth media for MSC culture because it provides growth factors, nutrients and extracellular matrix proteins which support MSC cells in vitro.11,12 There is also evidence that serum may act as an antioxidant for cells.13 Despite its zoogenic content, animal serum has been used since the first isolation of MSCs and remains a prime component for their culture and differentiation. Different methods for reducing animal antigens in fetal calf serum (FCS) have been suggested but none alleviate 100% percent risks,14 which leads to current researches for the development of substitute culture conditions, and a move towards the possible use of cheap, readily available as well as potentiate serum from other sources. The purpose of this study was to use three different types of serum for the culture and proliferation of equine adipose tissue derived mesenchymal stem cells (hrs-AT-MSC).
... 3,4 MSCs have been defined as non-hematopoietic progenitors able to self-renew, 5 migrate to a site of injury, 6,7 differentiate into mesodermal lineages, 8 modulate the immune response, 9,10 and secrete anti-inflammatory molecules. 11,12 These cells can also be easily isolated from different animal species 13 and preserved ex vivo, and they are considered safe because of their low immunogenicity after transplantation. 14,15 For the last decade, MSCs have been considered advanced medicinal therapy (AMT) and, therefore, compared with drugs; however, their mechanism of action (MoA) and tissue distribution in several target diseases are still unexplored and not completely understood. ...
Recently, mesenchymal stromal stem cells (MSCs) have been proposed as therapeutic agents because of their promising preclinical features and good safety profile. However, their introduction into clinical practice has been associated with a suboptimal therapeutic profile. In this review, we address the biodistribution of MSCs in preclinical studies with a focus on the current understanding of the pharmacodynamics (PD) and pharmacokinetics (PK) of MSCs as key aspects to overcome unsatisfactory clinical benefits of MSC application. Beginning with evidence of MSC biodistribution and highlighting PK and PD factors, a new PK-PD model is also proposed. According to this theory, MSCs and their released factors are key players in PK, and the efficacy biomarkers are considered relevant for PD in more predictive preclinical investigations. Accounting for the PK-PD relationship in MSC translational research and proposing new models combined with better biodistribution studies could allow realization of the promise of more robust MSC clinical translation.
... With the advancement in the field of cell culture technology, many workers have also reported successful culture and transplantation of osteoprogenitor cells derived from embryonic and mesenchymal stem cells at the fracture site for augmenting the healing process in different studies. 13,14,22 Protocol for superovulation, embryo recovery and collection, rabbit embryonic stem cell derivation and propagation and expression of the pluripotent marker in cultured rabbit embryonic stem cells performed in this study was found easy and very useful method. Similarly, the protocol for isolation of fetal skin fibroblast cells and their subculture used in this study were also found as very useful technique. ...
Role of Embryonic stem cell in osteogenesis
... With the advancement in the field of cell culture technology, many workers have also reported successful culture and transplantation of osteoprogenitor cells derived from embryonic and mesenchymal stem cells at the fracture site for augmenting the healing process in different studies. 13,14,22 Protocol for superovulation, embryo recovery and collection, rabbit embryonic stem cell derivation and propagation and expression of the pluripotent marker in cultured rabbit embryonic stem cells performed in this study was found easy and very useful method. Similarly, the protocol for isolation of fetal skin fibroblast cells and their subculture used in this study were also found as very useful technique. ...
... Intramuscular administration of xylazine (5mg/kg body weight) and ketamine (50mg/ kg body weight) provided safe and satisfactory anaesthesia for creation of corneal defect and its subsequent reconstruction with different biomaterial scaffolds in rabbits. The same anaesthetic protocol was reported to be effective for the collection of bone marrow from the iliac crest of rabbits (Maiti et al. 2013). ...
Clinically healthy adult New Zealand white rabbits (27) of either sex, were randomly divided into three groups (A, B and C) having 9 animals each. Porcine cornea was made acellular by treating it with 1% sodium dodecyl sulphate (SDS). Rabbit bone marrow derived mesenchymal stem cells were seeded over this acellular matrix. A 5 mm diameter lamellar keratectomy wound was created over the peripheral cornea of rabbits in all the 3 groups. In group A, the corneal defect was managed by simple tarsorrhaphy without any graft and is treated as control. In group B, defect was repaired with decellularized porcine cornea and in group C, corneal defect was repaired with r-MSC seeded decellularized cornea. On the basis of clinical, pathological and scanning electron microscopic examinations, mesenchymal stem cell seeded corneal scaffold showed better healing and vision when compared to nonseeded scaffolds. Cell seeded corneal matrix was found to be an alternative to conventional means of surgical management of corneal ulcer.
... All these research findings suggest the non-irreversible nature of the differentiation of various cell populations otherwise seen as an end point of variety of lineages. Protocol for culture of mesenchymal stem cells derived from bone marrow of rabbit and other animal species was reported [21,22]. Stem cell seeded biomaterials are now used routinely for different surgical affections [23,24]. ...
Mesenchymal stem cells derived from bone marrow have significant capacity for attachment, expansion and multilineage differentiation. This property has been widely used in the clinical practice for treatment of variety of diseases. Stem cell collection and culturing techniques varies for different species. Time taken for the primary cell culture to attain confluency depends on the species from which bone marrow is isolated. This paper describes an easy technique for the isolation and differentiation of mesenchymal stem cells derived from rat bone marrow (r-MSCs) on the basis of their density gradients and plastic adherence property along with the multilineage differentiation potential. After isolation, characterization was performed morphologically by examination under phase contrast microscope and viability detected by cell viability assay. Spindle shaped morphology was exhibited by the attached nucleated cells. Their multilineage differentiation to osteoblast, adipocytes, osteoblasts and chondroblasts was examined with respective induction medium and confirmed by oil red O, alizarin red and Alcian blue stains respectively. The r-MSCs were cultured and passaged for their expansion and were maintained in suitable environment for long time and can be used for cell based regenerative medicine.
... Isolation and culture of rBMSC was performed as described earlier [17] . The rBMSCs at passage-3 were used for this experiment. ...
Bone marrow derived mesenchymal stem cells (BMSC) represent an attractive cell population for tissue engineering purpose. The objective of this study was to determine whether the addition of recombinant human bone morphogenetic protein (rhBMP-2) and insulin-like growth factor (IGF-1) to a silica-coated calcium hydroxyapatite (HASi) - rabbit bone marrow derived mesenchymal stem cell (rBMSC) construct promoted bone healing in a large segmental bone defect beyond standard critical -size radial defects (15mm) in rabbits. An extensively large 30mm long radial ostectomy was performed unilaterally in thirty rabbits divided equally in five groups. Defects were filled with a HASi scaffold only (group B); HASi scaffold seeded with rBMSC (group C); HASi scaffold seeded with rBMSC along with rhBMP-2 and IGF-1 in groups D and E respectively. The same number of rBMSC (five million cells) and concentration of growth factors rhBMP-2 (50µg) and IGF-1 (50µg) was again injected at the site of bone defect after 15 days of surgery in their respective groups. An empty defect served as the control group (group A). Radiographically, bone healing was evaluated at 7, 15, 30, 45, 60 and 90 days post implantation. Histological qualitative analysis with microCT (µ-CT), haematoxylin and eosin (H & E) and Masson’s trichrome staining were performed 90 days after implantation. All rhBMP-2-added constructs induced the formation of well-differentiated mineralized woven bone surrounding the HASi scaffolds and bridging bone/implant interfaces as early as eight weeks after surgery. Bone regeneration appeared to develop earlier with the rhBMP-2 constructs than with the IGF-1 added construct. Constructs without any rhBMP-2 or IGF-1 showed osteoconductive properties limited to the bone junctions without bone ingrowths within the implantation site. In conclusion, the addition of rhBMP-2 to a HASi scaffold could promote bone generation in a large critical-size-defect.
... After attaining 70-80% confluence, culture was passage with Trypsin-EDTA solution (0.25% Trypsin and 1Mm EDTA (Gibco)). Third passage cells were used for further differentiation studies [14]. ...
