Composition Comprising Separated or Proliferated Cells from Umbilical Cord Blood for Treating Developmental and/or Chronic Lung Disease

Abstract

The present invention relates to a composition comprising separated or proliferated cells from umbilical cord blood for treating developmental and / or chronic lung disease, more precisely a composition containing separated or proliferated cells from umbilical cord blood for intratracheal administration for treating developmental and / or chronic lung disease. The cells of the composition of the present invention have excellent activities of proliferation and differentiation, so that the extraction, separation and selection of a tissue donor are all very easy. Besides, the composition of the invention can be very effectively used for the treatment of developmental and / or chronic diseases by intratracheal administration.

Description

TECHNICAL FIELD[0001]The present invention relates to a composition comprising separated or proliferated cells from umbilical cord blood for treating developmental and / or chronic lung disease, more precisely a composition containing separated or proliferated cells from umbilical cord blood for intratracheal administration for treating developmental and / or chronic lung disease.BACKGROUND ART[0002]The developmental and / or chronic lung disease includes adult chronic obstructive lung disease (COPD) such as cystic fibrosis, emphysema, and bronchopulmonary dysplasia of infant or premature baby, etc. The seriousness of these diseases is that there is no specific treatment modality even though the severeness and chronicity of these diseases cannot be disregarded. The bronchopulmonary dysplasia is a chronic lung disease developed during the treatment of respiratory failure in a newborn infant or a premature baby. Recent progress of treating a premature baby increases the incidence of this un...

AI Technical Summary

Benefits of technology

[0019]Umbilical cord blood, the origin of therapeutic cells of the invention, is defined as blood taken from umbilical vein connecting placenta and a fetus. Umbilical cord blood is a natural by-product of childbirth. The umbilical cord blood is much easier to obtain than general mesenchymal tissue like bone marrow requiring several steps of operation and it is also very easy to find a donor because umbilical cord blood deposit industry is developing steadily and infra has already been established. In addition, umbilical cord blood-originated cells are the one that does not express histocompatibility antigen HLA-DR (class II) which is the major cause of rejection after tissue- or organ transplantation. Thus, these cells can minimize the immune response according to transplantation, for example rejection against transplanted tissue or organ, suggesting that autologous or homologous umbilical cord blood transplantation is very useful and effective.

[0022]A composition of the present invention contains one or more cells selected from a group consisting of monocytes such as autologous hematopoietic stem cells and mesenchymal stem cells derived from the umbilical cord blood, mesenchymal stem cells derived from the umbilical cord blood, and mesenchymal stem cells sub-cultured and amplified from the mesenchymal stem cells. The mesenchymal stem cells derived from the umbilical cord blood are multipotent, unlike the typical stromal cells of bone marrow, suggesting that they are able to be differentiated into mesenchymal tissues such as bone, cartilage, adipose tissue, muscle, tendon, etc., under a proper condition. Umbilical cord blood derived mesenchymal stem cells have ability of self-renewal, suggesting that they are capable of proliferating under a proper condition, and might exhibit anti-inflammation activity when transplanted. Compared with those cells originated from mesenchymal stem cells derived from general mesenchymal tissues such as bone marrow, muscle and skin, these more primitive cells have far much excellent cell proliferation, and differentiation, and regulation substance secreting capacity.

[0025]The cell suspension is distributed into glass- or plastic ampoules for deep freezing, and then the ampoules are sealed and put in a deep freezer programmed to proper temperature. At this time, it is preferred to use a freeze-program allowing −1° C. / min of temperature change in order to reduce cell damage during thawing.

[0028]According to the present invention, cell concentration is 1.0×105-1.0×109 cells / Ml, and 1.0×106-1.0×108 cells / Ml is more preferred and 1.0×107 cells / Ml is most preferred. A composition of the present invention is preferably intratracheally-administered as close to lung tissues as possible, to increase therapeutic effect by improved accessibility, compared with the conventional cell transplantation using intravenous injection.

[0029]The present inventors centrifuged the extracted umbilical cord blood to separate monocytes, then the separated cells were cultured with a proper density in a culture vessel. When the cells were grown to proper density, sub-cultures were performed. The present inventors established a bronchopulmonary dysplasia model in neonatal rats by administering high-concentrated oxygen continuously from the birth. Bronchoalveolar lavage were obtained and the lungs were extracted from the animal model and stained. As a result, rats with bronchopulmonary dysplasia exhibited increased respiratory rate, poor weight gain, and chronic inflammatory reactions with monocytic infiltration and fibrosis with over-proliferated interstitial fibroblasts in the lung (see FIG. 1-FIG. 3). Besides, radial alveolar count (RAC) representing the number of alveoli was significantly decreased mean linear intercept (MLI) representing the size of alveoli was remarkably increased, and resultantly the ratio of RAC to MLI, an alveolar development index, was significantly reduced, compared with that in the wild type normal rat.

[0032]The cell culture medium of the present invention can additionally include one or more subsidiary components, for example, fetal bovine serum, horse serum or human serum, antibiotics such as Penicillin G or streptomycin sulfate and antifungal agent such as amphotericin B, gentamycin or nystatin to prevent microorganism contamination.

Problems solved by technology

The seriousness of these diseases is that there is no specific treatment modality even though the severeness and chronicity of these diseases cannot be disregarded.

Not only this disease is regarded as a major cause of death of a newborn infant or a premature baby, but also among survivors, long-term hospitalization is required and serious side effects including pulmonary hypertension must be a worry.

However, steroid treatment is now limited in relation to the recent reports that the use of steroid might be responsible for later abnormal neurodevelopmental prognosis, especially increase in cerebral palsy, etc (Committee on Fetus and Newborn, Pediatrics, 109: 330-8, 2002).

However, the transplantation of embryonic stem cells which have excellent differentiation potential has serious technical and ethical problems, that is generation of uncontrollable teratoma, developmental problems caused by genomic imprinting, and questions of moral and ethics.

Therefore, the application of embryonic stem cells seems to be limited and instead adult stem cells have been moved into the center of our interest.

Nat Immunol., 3:329-333, 2002), and bone marrow extraction itself is distressing, which is disadvantage for actual clinical application.

At first, it was a common belief that the number of mesenchymal cells to be taken from umbilical cord blood was small and the proliferation of them was very difficult.

However, there has been no attempt to apply umbilical cord blood derived stem cell transplantation to developmental and / or chronic lung disease.

However, there have been no descriptions on the therapeutic effect of umbilical cord blood-derived stem cell transplantation in developmental and / or chronic lung disease.

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