Implementation of biotechnologies applying cells and tissue cryoconservation method into clinical practice

Long and time consuming process of scientific research starting from 1971 and till nowadays, performed by Lobyntseva Galina Stepanovna, researcher of the Institute of Cellular Therapy, candidate of medical sciences, and senior researcher by specialty “Cryobiology”, preceded introduction of biotechnologies with application of cells and tissues cryoconservation method into clinical practice.

As a result of these studies scientific approach in medicine named “cellular and tissue therapy” was established.

At the first stages of the research the objectives of fundamental exploration of cryodamage and cryoprotection mechanisms of biological objects, possibilities to manage them and creation of scientifically justified cryoconservation regiments that preserve morphological and functional values of cells and tissues were set up.

The cells from blood and bone marrow were the most convenient objects for experiments. The results of research were formalized in the form of scientific reports, publications and invention applications. A number of certificates of recognition was received proving priority and innovation of given research.

The research of cryoconservation of embryonic hemopoietic human cells started by G.S.Lobyntseva in 1980 was specifically interesting. Application of the mathematic modeling method in experiments put special demands not only to equipment that was custom-designed in the Model shop of the Institute of Cryobiology and Cryomedicine Problems of the Academy of Sciences of USSR, but to tests of cells’ biological value assessment after cryoconservation. The method of cultivation in semi-liquid agar media can be mentioned as the most accurate and reliable method of hemapoietic cells-precursors value assessment.

G.S.Lobyntseva has mastered this method in Moscow Oncological Institute under instruction of S.I.Chereshkov, candidate of biological sciences, and L.A.Kharlamova, candidate of biological sciences, who worked earlier in the leading laboratory on cultivation of hemapoietic cells of I.L.Tchertkov, doctor of biological sciences.

Such approach allowed revealing patterns of cryoconservation factors interaction and to formulate assumptions about elimination of main cryodestruction factors by manipulations with the rates of temperature decrease at the different sectors of temperature scale, by cryoprotectors concentrations and temperature halting that allow changing the character of crystals formation in object under freezing. As a result, the program of cryoconservation of hemapoietic cells of embryonic liver with 5% concentration of cryoprotector dimethyl sulfoxide has been developed and protected by the patent. This excluded the procedure of washing from DMSO before administration of cells to recipient, which complicated all the process and led to cell loss.

In 1980 a complex scientific program of the Academy of Sciences of USSR and Academy of Medical Sciences of USSR “Fundamental sciences for medicine”, All-union program 0.69.14 “To develop and to introduce the methods, technical means and new technological processes of cold effect on biological objects into different areas of national economy” was established.

Scientific theme “To develop method of cryoconservation of human embryonic liver hemapoietic cells (in -196º C)” that was completed with 5-year clinical trial of the preparation “Cryoconserved hemapoietic cells of human embryonic liver”, approval of methodic recommendations “Clinical application of cryoconserved hemapoietic cells of human embryonic liver” (Kiev, 1991, re-approved in 1996) and “Technological regulations for receipt and cryoconservation of hemapoietic cells from human embryonic liver for clinical application” (Kiev, 1995) was executed in the framework of objective of the State Committee for science and technique under the leadership of G.S.Lobyntseva (who was the principal investigator).

In 1990 G.S.Lobyntseva defended doctor’s thesis “Cryoconservation of hemapoietic cells from embryonic liver” in the Institute of Cryobiology Problems of the Academy of Sciences of Ukraine in unanimous voting of Academic Council members, where theoretic, experimental and clinical results of the research performed on hematopoietic embryonic cells of humans and animals (rabbits, calves, pigs) were reported.

At that time Ukraine did not have its own Highest Attestation Commission, that is why the dissertation was sent to the Highest Attestation Commission of the USSR, where is was reviewed at the commission on veterinary without a single specialist neither in hematology nor in cryobiology. Commission members have not understood all the importance of the work that opened track for whole scientific direction in medicine of “cellular therapy” and pronounced the verdict: the theme is narrow, non-perspective and proposed to take back the dissertation for working it over. However, in 1992 G.S.Lobyntseva was awarded with State Prize of Ukraine in the area of science and technology for the series of works “Establishment of scientific background and methods of cryoconservation of cell suspensions and their application in medicine”. Preparations, produced by G.S.Lobyntseva technology, are known both in our country and abroad. A contract agreement between Institute of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine and American company was being performed over 3 years (1995-1997) to deliver experimental samples of embryonic hemapoietic and nervous cells.

Over the time passed after receipt of the documents authorizing clinical application of the method hemapoietic cells preparations were administered in very different clinics of Ukraine and Russia, results of cells application in oncological, hematological, urological, surgical, gerontological and other practice have been accumulated and summarized.

In 2004 G.S.Lobyntseva and co-authors published a monography “Stem embryonic hemapoietic human cells (theory and clinical practices)" where summarized authors’ experience in application of cryoconserved stem embryonic hemapoietic cells in clinical practice was stated and possible mechanisms of positive cells action in different pathologic organism conditions were analyzed.

All the huge scientific experience of G.S.Lobyntseva, accumulated in the area of cryobiological research at present time is directed towards provision of quality work of the First in Ukraine Cryobank of umbilical blood stem cells established on the basis of the Institute of Cellular Therapy.

Developed technology of mononuclei cryoconservation allows saving all cell population practically without losses, providing guarantee of successful clinical application of stem cells prepared and stored in liquid nitrogen.