Stem cells exist and can do a lot. For several decades, bone marrow and umbilical cord blood transplants containing haematopoietic stem cells have been used in the treatment of blood diseases. Genetic modifications (gene therapy) of such cells help to cure complex immunodeficiencies and severe anaemias. The limbal stem cells taken from the eye and properly multiplied can regenerate the damaged cornea, and the epidermal stem cells help in the treatment of severe burns and some hereditary, severe skin diseases. Promising experimental research is under way on other uses of stem cells. However, these cells are appropriately selected, having real ability to differentiate into specialized cells whose malfunction is the cause of the disease. Therapeutic applications of stem cells are apparently limited to date. Meanwhile, the Internet is full of advertisements for supposedly miraculous treatments for almost any disease. Stem cells have become a modern synonym of the Holy Grail. A wonderful dish, transforming every drink into elixir of health, youth and long life. Stem cells from a single source, e.g., umbilical cord blood, or so-called cells, although without proven properties of stem cells, are offered in commercial private clinics as a panacea for autism, cerebral palsy, spina bifida, eye diseases, amyotrophic lateral sclerosis and dozens other disorders. Without justification for their action in these diseases, without convincing evidence of safety, but for a high fee. This article discusses stem cells and misunderstandings about including any cells among them. It draws attention to the real possibilities and confirmed uses of stem cells and presents the problems, doubts and dangers for patients associated with commercial offers of treatments using “stem” cells. The author cites the positions of scientific institutions and societies warning against premature commercialization of unjustified and potentially dangerous therapies.

The aim of the study was to compare the effects of corneal healing in case of application of stem cells in various forms, in relation to the antibiotic-assisted procedures. Rabbits were divided into 4 groups in the first stage of the experiment. Group 0 (negative control group) was not subjected to any actions, which would cause damage to the cornea. The remaining three groups had their cornea damaged. Group 1 (positive control group) – no drugs were administered during the experiment. Rabbits in group 2 were administered with ointment containing stem cells to the lesion, while group 3 – with ofloxacinum. The stem cells were administered during the first five days, twice a day, onto the corneal surface. The further course of the experiment consisted of observing the rate of healing of the injured cornea and assessment of its transparency, size of lesion, hyperaemia, eyelid spasm and outflow from the conjunctival sac after 5, 10 and 20 days.

In the second stage the animals were euthanised after clinical examination on the twentieth day of the experiment, in order to analyse the corneal reparative processes on the same day. The studies revealed that the application of antlerogenic stem cells had a positive effect on the healing process of corneal defects. The application thereof not only shortened the healing time, but also weakened or arrested the development of side effects. The results have demonstrated that the epithelial proliferation in each group was different. The longest was maintained in the group with stem cells, the shortest – in the group with chemotherapeutics. The use of antlerogenic stem cells had a positive effect on the healing process of corneal lesions. The use of stem cells helped to maintain high transparency of the cornea.

Cellular therapy, as a part of regenerative medicine, implies to the treatment of human disorders with cells as a medical product, so called – “living drugs”. Usually such therapy is applied when other alternative efficient pharmacological therapies are not available. Stem cells of different origin: 1) tissue specific e.g. hematopethic, epithelial, neuronal, limbal; 2) mesenchymal stem cells (MSC) harvested from variety of tissues; 3) pluripotent stem cells: embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) – serve as a source of cells for regenerative medicine application, depending upon disease and application requirements. Currently MSC are the type of stem cells that are most frequently used in registered regenerative medicine clinical trials. In this paper we provide the information on the application of cell therapy in orthopedics, hematology, ophthalmology, dermatology, gastrology and neurology. The influence of origin of MSCs and iPSCs on their mode of action as therapeutic, regenerative agents are discussed. Advantages and disadvantages of application of different cell types for cell therapy are underlined. Last, but not least current low regulations in Poland and requirements of European regulatory bodies for cell therapy are pointed out and discussed.

One of the most common reasons for horse lameness is subchondral bone cysts (SBCs), which are especially evident in young horse athletes. It is believed that SBC development is strongly associated with an individual’s bone growth and/or bone microstructure impairment. Current methods of SBC treatment include pharmacological treatment or surgical procedures which may allow the bone within the cyst to rebuild and be restored to properly developed bone tissue. Thus, we propose filling the SBCs with a 3D complex of alginate hydrogel and autologous adipose derived mesenchymal stem cells (ASCs). We have observed at the in vitro level, that this hydrogel complex induces osteogenic and chondrogenic differentiation potential through the upregulation of bone morphogenetic protein, osteopontin, collagen type I and aggrecan mRNA levels. Moreover, we detected the creation of a 3D extracellular matrix (EM). To investigate the complex in vivo, we chose 8 horses of varying age suffering from SBC, which resulted in lameness, to undergo experimental surgery. We documented the horses’ clinical appearance, lameness and radiographic appearance, to determine that there was clinical improvement in 87.75% of the patients (n=7, out of 8 horses) 6 months postoperatively and 100% (n=8, out of 8 horses) a year after surgery. These results are promising for the potential of this procedure to become the standard in SBC treatment.

Novel tendinopathy treatment protocols should be assessed for safety. The goal of this work was to compare differences in selected systemic inflammatory marker concentrations after two treatment protocols for collagenase induced Achilles tendinopathy in sheep. 14 sheep (aged 5 and 6 years, Polish Mountain Sheep breed, weight 60-70kg) were injected with bacterial collagenase type 1A-S (Clostridium histolyticum, C-5894, Sigma Aldrich, Poznań, Poland) bilaterally to Achilles tendons. Subsequently, the animals were injected with Platelet Rich Plasma (7 sheep) or Adipose Derived Stem Cells (7 sheep) to induced tendinopathy foci. Left limbs of all sheep were additionally treated with Radial Pressure Wave Therapy (RPWT) focused above the tendinopathy origins. Treatment progress was controlled by ultrasound scans, and tendon samples were taken on the 126th day of the experiment. Serum Amyloid A (SAA) concentration showed mild elevation before the experiment (2 sheep from group I, 4 sheep from group II) and two days after the intratendinous growth factors injection ( 4 sheep from group I, 3 sheep from group II) combined with RPWT (mean 22,63 mg/L and 53, 6 mg/L respectively). Haptoglobine (Hp) concentration increased from 0 to 0,01 g/L in 2 animals from group I two days after injection. These values declined to 0 during the course of the treatment. Fibrinogen (Fb) concentrations were within reference levels throughout the research, although mild elevation was observed before the treatment course in 6 sheep from group I and 1 sheep from group II. In conclusion, addition of RPWT to growth factors injections in the treatment of yatrogenic Achilles tendinopathy in sheep did not induce systemic inflammatory response.

Cell culture transplantation is very promising in the treatment of various diseases. Cells obtained from a number of sources have been analysed to provide a basis for further studies in the area of regenerative medicine. The objective of the study was to compare morphological and phenotypic changes in cat adipose tissue and bone marrow cell cultures from the first to fifth passages. Adipose tissue and bone marrow were used to obtain cell cultures (coming from 3 cats) using standard methods with own modification. Phenotype changes were monitored by CD-marker identification and CD pan-keratin. The cytogenetic analysis was performed on 50 metaphase plates of cell cultures from the first to fifth passage. Cytogenetic assays showed that the adipose tissue cell culture (ATCC) at all passages was more stable than the bone marrow cell culture (BMCC).

O b j e c t i v e: The main goal of our studies was to investigate the eff ect exerted by pulsed electromagnetic filed (PEMF) on adipocytokines secretion in cell culture supernatants from rat adipose derived stem cells (ADSCs) grown on varied energy-rich diet. Off spring and adult animals were randomly selected for two types of experimental diets: low (LF) or high fat (HF) diet for 7 weeks. After the diet period, serum glucose level was measured, ADSCs were isolated from adipose tissues from different locations. ADSCs from all experimental groups were exposed to PEMF, supernatants collected and adipokines level was determined.

R e s u l t s: HF diet feed in pups/adult animals elevated blood glucose level and increased the level of adiponectin (Apn) and leptin of both genders and age measured in serum. ADSCs cell cultures originated from female pups on LF diet and exposed to PEMF released large amounts of Apn. PEMF effect exerted on Apn release was also observed in ADSCs isolated from male pups HF diet. ADSCs from female pups on LF diet exposed to PEMF released smaller amounts of leptin in comparison to cell cultures without PEMF treatment. PEMF exposure of ADSCs cell cultures originated from female adults on LF diet decreased release of Apn, contrary adult male on LF diet ADSCs under PEMF treatment produced more leptin. PEMF treated male HF diet-originated ADSCs cultures released significantly more leptin than controls.

C o n c l u s i o n: Our results suggest that PEMF exposure is responsible for metabolic physiological balance effects obtained in ADSCs cultures originating from adult animals on HF diet.

Introduction: Uterine leiomyoma is the most widespread benign tumor affecting women of childbearing age. There are still gaps in the understanding of its pathogenesiss. Telocytes are unique cells described in greater than 50 different locations inside the human body. The functional relationship of cells could clarify the pathogenesis of leiomyomata. In the current study, we focused on the identification of telocytes in all regions of the human uterus to explain their involvement in leiomyoma development.

Materials and Methods: Tissue samples from a healthy and myomatous uterus were stained for c-kit, tryptase, CD34 and PDGFRα to identify telocytes. Routine histology was performed to analyze tissue morphology and collagen deposits.

Results: Telocytes were detected in the cervix, corpus of the uterus and leiomyoma. The density of telocytes in fibroid foci was reduced compared with normal myometrium.

Conclusions: Our results demonstrated the existence of telocytes in all parts of the human body affected and unaff ected by leiomyoma of the uterus. In addition, telocytes were also present in leiomyoma foci. Our results suggest that the reduced density of telocytes is important for the pathomechanisms of myometrial growth, demonstrating its value as a main component of the myomatous architecture.

Telomerase reverse transcriptase (TERT) vectors were transfected into bone marrow mesen- chymal stem cells (BMSCs) which were then cultured and selected to establish TERT-BMSC cell lines whilst sequencing BMSCs and TERT-BMSCs via transcriptome in this study to explore their regulatory mechanism and effect on osteogenic differentiation after TERT ectopic expres- sion in sheep BMSCs. After sequencing and analysing differential genes, PI3K/Akt signalling pathway related to osteogenic differentiation was investigated. Western blot was used before and after applying the PI3K/Akt signalling pathway inhibitor LY294002 to detect protein expression levels of AKT and p-AKT. On the twenty-first day of osteogenic differentiation, RT-qPCR and Western blot were used to detect mRNA and protein expression levels of RUNX2 and OPN and alizarin red staining was utilised to analyse calcium salt deposition. Results showed that pro- tein expression levels of AKT and p-AKT were significantly up-regulated, mRNA and protein expression levels of RUNX2 and OPN increased and calcium salt deposition increased after ectopic expression of TERT. After applying LY294002, the protein expression of AKT and p-AKT was down-regulated, mRNA and protein expression levels of RUNX2 and OPN were reduced and calcium salt deposition was reduced. These results confirmed the stable integration and expression of the exogenous TERT gene in BMSCs to promote the differentiation of BMSC osteoblasts, which may be mediated by the PI3K/Akt signalling pathway.

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apoptosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin-dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluorescence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly, cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

Knowledge of uterine fibroids has existed since the time of Hippocrates. However, there are still wide gaps in the understanding of its pathogenesis. No single theory explains the background of uterine fibroid pathology, which affects more than 50% of women worldwide. By contrast, a newly depicted cell type called telocytes was only recently identified in the past twenty years. Th ese cells have evoked ambivalent opinions in the scientific community. The unique features of telocytes coupled with experimental evidence by numerous researchers and our hypotheses and conceptions are discussed in this review. We emphasize the main telocyte interactions in the context of the uterine fibroid architecture. This review reveals the pivotal role of telocytes, describing their contacts with smooth muscle cells, fibroblasts, vessels and nerves, inflammatory cells and stem cells. Our data are based on the latest publications and our own results.