Dr Peter Eftimov
General information
Mesenchymal stem cells (MSC) are multipotent stromal cells that can differentiate into a variety of cell types, including osteoblasts, chondrocytes, and adipocytes. There are evidences that MSC can be differentiated into oligodendrocyte and dopaminergic neurons, with further clinical applications.
Because the cells, called MSCs by many labs today, can encompass multipotent cells derived from other non-marrow tissues, such as umbilical cord blood, adipose tissue, adult muscle, corneal stroma or the dental pulp of deciduous baby teeth, yet do not have the capacity to reconstitute an entire organ, the term Multipotent Stromal Cell has been proposed as a better replacement.
The International Society for Cellular Therapy has provided the following minimum criteria for defining multipotent human mesenchymal stromal cells:
1) plastic-adherent under standard culture conditions;
2) positive for expression of CD105, CD73, and CD90, and absent for expression of hematopoietic cell surface markers CD34, CD45, CD11a, CD19, and HLA-DR;
3) under specific stimulus, cells should differentiate into osteocytes, adipocytes, and chondrocytes in vitro.
Case reports
Treatment of severe osteoarthritis in 12 years old German sheperd dog vie celluar-substitution therapy with Mesenchymal Stem Cells
Duc, 12 years old German Shepard was presented at the clinic with progressive lameness of the pelvic limbs, which deteriorates in cold days, exercise intolerance and signs of severe pain. He was treated for over a year with NSAID (including selective COX-2 inhibitors), combined with food supplements. The treatment didn’t succeed to alleviate symptoms – in fact, when presented at clinic the patient barely could move his legs and have impaired urinary function.
After confirming the diagnosis with a radiographic examination (fig.1), and taking into account the age of the animal, we decided to attempt cellular-based treatment using adipose derived MSCs, rather than total hip replacement.
Biopsy of approximately 20 grams subcutaneous fat was taken and MSCs (CD44+, CD90+, CD105+) were derived, following standard protocol. After two passages they reached target concentration of 3.106cells/ml and were applied intra-articulary in both coxofemoral joints. Pain alleviation with sub second improvement of the gait was observed 6 days after treatment, and 4 weeks after application of the MSC the dog tolerates long walks (over 40 minutes) and doesn’t show any signs of pain.
Six months after the procedure, Duc visited us again this time with progressive osteoarthritic changes in both elbows and cauda equina syndrome (fig. 2). Another course of cell-based treatment was suggested – via intravenous infusion of stem cells, but the owner preferred topical treatment with hyaluronic acid. It was applied in both elbow joints, but after mild improvement Duc condition deteriorates (lameness with thoracic limbs, urinary incontinention, severe pain and loss of appetite) and after another 3 months he was euthanized.
It is remarkable that he never experienced pain in hip joints after MSC treatment. That fact and the fast recovery after first intervention were highly appreciated by the owner, but the relatively high cost of the cellular therapy, prevented him for taking the decision for another procedure
Treatment of chronic kidney disease (CKD) in a dog and in a cat
1. Treatment of 5 years old male dog with CKD
Lucky is a 5 years old miniature Poodle was presented in deteriorated condition with ongoing conservative treatment of CKD stage 3 (according to IRIS classification) After ultrasound examination we decided to include him in the program for MSC treatment.
Biopsy was performed and MSC were isolated and cultured to the therapeutic concentration (2*106 cells/ml).
Due to his severe condition cells were applied intravenously (to avoid general anesthesia risk). After initial improvement, Lucky deteriorated and discontinued food intake. Another procedure was made and this time cells were administered intra-renaly under the ultrasound guidance. Prior to that, he was subjected to a peritoneal dialysis, in order to diminish the levels of toxic metabolites. Unfortunately, no definitive improvement was observed and shortly after the procedure, the owners took decision to euthanize Lucky
2. Treatment of 14 years old male cat with CKD
Macho, 14 years old cat, was presented with typical signs of CKD (weight loss, decreasing of the appetite, polydipsia and polyuria). He was subjected to a peritoneal dialysis one year ago, after deterioration of his condition and since then was on a standard protocol for treatment of CKD i.e., nutritional supplementation and special diet.
After initial examination ,a biopsy of subcutaneous adipose tissuewas taken, and MSC were isololated and prepared for intra-renal application,which was made under ultrasound guidance.The improvement was quick and stable- the cat regained his appetite, and the values of creatinine measured after the procedure continues to decrease.
We work on a second case of a cat with CKD now.
Treatment of n.radialis paralysis in a cat.
Naida was found on the street and brought to our clinic with a dull trauma of the abdomen, elbow bent low, curved dorsal paw and inability to stepping on his front right leg. Following the screening, we found lack of surface sensitivity on the rostral surface of the limb and lack of deep sensitivity. Our working diagnosis was trauma to the radial nerve. The condition is more common in dogs than in cats and is often the consequence of a hit by a car or falling from above. After considering other options as carpal arthrodesis, amputation or transposition of n.ulnaris we chose to start a procedure of MSC treatment.
Following the standard procedure, a biopsy from the abdominal adiposal tissue was taken and MSC were isolated and cultured. A highly concentrated stem cells isolate was injected into the perineural space of n. radialis.
Two weeks after implantation of mesenchymal stem cells we observed (although minimal) surface and deep sensation in the affected limb. Najda began to flex her elbow, indicating recovery of n. musculocutaneus.
