Trans-Technologies's programs, 2004/2007 years.
Trans-Technologies Ltd was established in 2002 as a private company for mesenchymal stem cells biology study. Our company investigates one of the most promising areas of contemporary medicine – the possibility of the stem cells therapy. We have two separate programs – commercial program (personal storage of cord blood cells) and scientific program (the biology of mesenchymal stem cells (MSC), preclinical studies of MSC-based therapy and clinical studies of auto MSC applications).
Commercial program:
Our cord blood program was initiated in 2005. Trans-Technologies is one of 12 organizations that are officially allowed to store cord blood cells at the Russian Federation, and we are the first St-Petersburg company to do that.
Scientific program:
1) Stem cell biology.
The area of our interests lies in the investigation of adult stem cells properties, to be exact, the MSC differentiation, secreting profile and migration ability. By the present time we have developed our own protocols (patent pending) for human and rat MSC isolation from the bone marrow.
In the frame of this part of scientific program we evaluate the differentiation potential of rat bone marrow MCS in vitro.
2) Preclinical studies of MSC-based therapy.
Mesenchymal stem cells (MSCs)-based therapy is a promising modern approach to the treatment of various pathologies. Means of MSCs transplantation are being widely discussed. MSCs can be transplanted directly into the damaged region, border zone of injury, or intravenously. We characterized the MSCs distribution in inflamed tissue (prostate) after various ways of transplantation. We found that MSCs actively migrate to the area of acute inflammation in the tissue stroma. We consider intravenous transplantation to be preferable as the cells injected into the vein were distributed diffusely and uniformly around the site of injury, whereas the MSCs transplanted directly to the damaged tissue were located as a compact group. Moreover, the intravenous injection is less traumatic, while the direct transplantation to inflamed tissue can cause an additional injury.
Publication: http://www.abstractsonline.com/
In experimental cardiology, we determined optimal time frame for the mesenchymal stem cells (MSCs)-based cell therapy approach. Experimental infarction operation (coronary artery occlusion) was performed on inbred rats. So our model was singeneic and correspond to auto MSCs therapy in humans. We demonstrated, that upon intravenous administration, MSCs migrate to the infarcted heart and promote cardiac repair (fig. 1, 2, 3) and vascularity restore (fig. 4). We detected, that the effect of the MSCs transplantation depends on the time of the injection. The data suggest that the best therapeutic effect is obtained after MSCs injection during the first week after the infarction.
Publications:
Krugliakov PV, Sokolova IB, Amineva KhK, Nekrasova NN, Viide SV, Cherednichenko NV, Zaritskii AIu, Semerin EN, Kisliakova TV, Polyntsev DG. The influence of mesenchymal stem cell transplantation time on myocardial reparation in rat experimental heart failure. Tsitologiia. 2005;47(5):404-16. Russian.
Krugliakov PV, Sokolova IB, Amineva KhK, Nekrasova NN, Viide SV, Cherednichenko NN, Zaritskii AIu, Semernin EN, Kisliakova TV, Polyntsev DG. Mesenchymal stem cell transplantation for myocardial reparation of rat experimental heart failure. Tsitologiia. 2004;46(12):1043-54. Russian.
Fig. 1. MSC are able to migrate into infarcted myocardium. 21-st day after infarction. MSC labeled with PKH26 (red), scar tissue (arrows)
Fig. 2. Histological changes in the hearts of control animals (A) and animals that received cell therapy (B) after myocardial injury; Hematoxilin-eosin, 10x. A. Forty-two days after operation. Scar area; B. Forty-two days after operation. Scar area, cardiomyocyte layer in the injured area.
Fig. 3. A. Labeled cardiomyocytes in boarder area of scar tissue (arrows). B. Connexin 43/PKH26 double positive cells.
Fig. 4. Vascularity of scar tissue. MSCs treatment for myocardium infarction led to vessels formation/survival in scar tissue.
In the experimental traumatology (bone defects), we used MSC to improve the reparation of damaged bone tissues (fig. 5). One aspect of the tissue engineering is concerned with the formation of three-dimensional tissue substitutes by culturing cells on natural or synthetic polymer scaffolds. In our studies, demineralised bone matrixes (DMBs) from parietal rat bones were used as bone substitutes. DMBs were loaded with the singenic MSCs (fig. 6). Histological examination showed obvious de novo bone formation and angiogeneic activity in the place of the osseous defect when the MSC-loaded matrixes were implanted. The MSC-loaded rat DBM implantation resulted in lamellar bone formation. This bone was at least partly united with the recipient bone (fig. 7).
Fig. 5. Experimental model of bone tissue trauma. We surgically eliminated a part of the cranium bone (circle). The defect square was 0.25 sm2. Than defect was replaced with control or experimental graft.
Fig. 6. Bone matrix seeded with MSCs (arrows).
Fig. 7. 119 days after bone injury. Rat demineralized bone matrix (A,B). Rat demineralized bone matrix seeded with singenic MSC (C,D). The contact of host bone (yellow arrows) and graft (red arrows) (A, C), graft area (B,D). We detected survived graft integrated to the host cranium.
We detected an immunosupressive property of the MSC in experiments with xenogenic bone matrixes (DMBs) from chicken bones. The MSC suppressed post-implantation inflammation and survival of the implantat in case of the MSC-loaded chicken DMBs (fig. 8).
Fig. 8. 119 days after surgical bone injury. Chicken demineralized bone matrix (A,B). Chicken demineralized bone matrix seeded with singenic MSC(C,D). The contact of host bone (yellow arrows) and graft (red arrows) (A, C), graft area (B,D). We detected total graft resorbtion in control group and survived graft with osteogenesis nidus in case of MSC treatment.
Publications: http://www.abstractsonline.com/
Krugliakov PV, Sokolova IB, Zin'kova NN, Viide SV, Cherednichenko NN, Kisliakova TV, Polyntsev DG. The influence of mesenchymal stem cells on bone tissue regeneration upon implantation of demineralized bone matrix. Tsitologiia. 2005;47(6):466-77. Russian.
In order to explore the effect of MSCs transplantation on a damaged brain, we carried out a set of experiments using the pre-clinical stroke model (middle cerebral artery occlusion method). We demonstrated that the intravenously administrated MSCs significantly improve recovery after the stroke. Also, we detected that the MSCs substantially reduce the lesion size (fig. 9), and improve vascularity. Moreover, a behavioral recovery was observed.
Fig. 9. Injury area in control and experimental animals. 6 weeks after experimental stroke.
Publications:
There are going to be opened:
MSCs-based therapy of diabetes type 2.
MSCs-based therapy of cartilage trauma.
MSCs-based therapy of brain trauma.
3) Clinical studies of MSC applications.
Clinical trials of the mesenchymal stem cells - based approach were started in 2003. We have a large experience with the human MSC transplantation. About 200 patients received the MSC – based therapy within our clinical program. All our clinical trials are ethical comities approved.
Current clinical protocols are:
1) The clinical protocol of autologous mesenchymal stem cells application for the treatment of patients with a chronic ischemic disease.
In this study, we detected highly raised physical training tolerance restoration and quality of life restoration. PET data showed metabolism changes in the scar tissue up to the level of the undamaged myocardium.
2) The clinical protocol of autologous mesenchymal stem cells application for the treatment of patients with invasive bladder cancer.
This study has not detected a stimulation of the tumour growth. The patients stood radiotherapy and chemotherapy easily, without hematological complications.
3) The clinical protocol of autologous mesenchymal stem cells for application an optimization of reparative osteogenesis.
Usage of the auto MSC in bone growth stimulation by Ilisarov technique led to the reduction of time need for the bone growth at least in 1,5 times.
Auto MSC – based treatment of long bone non-unions. Average periods of an external fixation in cases of standard postoperative periods were 3,5 months, with full restoration of weight-bearing, that is 2 times more quickly compared with not MSC seeded implants (fig. 10). X-ray and computing tomography data showed the areas of ossification in implants in all cases (fig. 11).
 |  |  |
Fig. 11. Computing tomography of bone formation after graft seeded with MSCs transplantation (arrows). A,C – before transplantation, B,E –after graft transplantation, C,F – 3 months after graft transplantation.
4)The clinical protocol of hematopoietic and non-hematopoietic (mesenchymal) stem cells co-transplantation for oncohematological diseases
In this protocol we used the allogenic MSC. The main idea of the project was to reduce GVHD after the allogeneic bone marrow transplantation. We detected the reduction of GVHD signs after the allo-MSC treatment.
5)The clinical protocol of autologous mesenchymal stem cells application for the treatment of patients with psychiatric disorders and brain trauma.
Transplantation of the cell material was carried out intravascularly or intracerebrally; in some cases both the variants were used simultaneously. Distinct improvement in the state of the patients' mental health was observed after the transplantation. It was demonstrated that in the first half of the year after the autologous MSC neurotransplantation, patients' mental health showed distinct improvement (p<0.02). Moreover, positive changes were detected in the clinical state of the patients: an emotional inadequacy disappeared, while the sensibility at play and elementary self-service skills showed up. After the appliance of MSC, distinct positive dynamic was observed in the overwhelming majority of the patients.
6)The clinical protocol of autologous mesenchymal stem cells application for the treatment of parodont diseases.
The first results of this study showed positive changes. In co-transplantation of auto MSC with hydroxyapatite (Cerasorb), a bone formation was observed in the parodont cavity (fig. 12).
Fig. 12. Bone density after MSC/Cerasorb treatment.
There are going to be opened:
The clinical protocol of application autologous MSC for the treatment of bone breaking.
The clinical protocol of application autologous MSC for the treatment of vascular diseases.
The clinical protocol of application autologous dermal equivalent based on MSC for the treatment burn.
Contacts:
Peter Kruglyakov
Stem cells laboratory, head
e-mail: pitkus@alkorbio.ru
tel.: +7-812-596-67-75
fax.: +7-812-596-67-80
Dmitriy Polintsev
General manager
e-mail: d.polyntsev@alkorbio.ru
tel.: +7-812-596-67-75
fax.: +7-812-596-67-80
