Human uterine cervical stem cell population and uses thereof

Abstract

The present invention relates to a method for isolating stem cells comprising preparing a cell suspension from uterine cervix tissue, to the stem cells isolated by said method, and to the conditioned medium obtained from the culture of said stem cells. The invention also encompasses the use of said stem cells or conditioned medium for treating or preventing cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders, as well as for cosmetic treatment.

Description

TECHNICAL FIELD OF THE INVENTION[0001]The present invention relates to a method for isolating stem cells comprising preparing a cell suspension from uterine cervix tissue, to the stem cells isolated by said method, and to the conditioned medium obtained from the culture of said stem cells. The invention also encompasses the use of said stem cells or conditioned medium for treating or preventing cancer, inflammatory diseases, autoimmune diseases chronic pathologies or infectious diseases, as well as for cosmetic treatment. Therefore, the present invention relates to the field of stem cells and the therapeutic or cosmetic use thereof.BACKGROUND ART[0002]A stem cell is characterized by its ability to self-renew and to differentiate along multiple lineage pathways. A particularly promising type of adult stem cells for therapeutic applications is the so-called mesenchymal stem cells (MSCs). MSCs, also defined as multipotent mesenchymal stromal cells, are a heterogeneous population of cel...

AI Technical Summary

Benefits of technology

[0014]The authors of the present invention have discovered that the uterine cervix tissue can be used as source of stem cells. The cells isolated from this tissue, called uterine cervical stem cells (UCESC), show a higher anti-inflammatory activity, anti-tumor capacity, antimicrobial activity and growth rate than other mesenchymal stem cells isolated from other tissues, and are capable of keeping their functionality and a stable karyotype for at least 10 cell passages. Additionally, the new source of stem cells allows the isolation of mesenchymal stem cells by a non-invasive and painless method since said tissue can be obtained just by exfoliating said organ during a routine gynaecological examination.

[0022]Steps (a)-(d) can be carried out by conventional techniques known by those skilled in the art. An additional advantage of this new source of stem cells is that it allows obtaining stem cells from the animal body in a non-invasive and painless way.

[0032]Once the cells proliferate, these may be maintained in culture in the same medium and under the same conditions until they reach the adequate confluence, typically, about 80% cell confluence, with replacement of the cell culture medium when necessary. After reaching the desired cell confluence, the cells can be expanded by means of consecutive passages using a detachment agent such as trypsin and seeding onto a bigger cell culture surface at the appropriate cell density (usually 2,000-10,000 cells / cm2). Thus, cells are then passaged at least two times in such medium without differentiating, while still retaining their developmental phenotype. More preferably, the cells can be passaged at least 10 times (e.g. at least 15 times or even at least 20 times) without losing developmental phenotype. Typically, the cells are plated at a desired density such as between about 100 cells / cm2 to about 100,000 cells / cm2 (such as about 500 cells / cm2 to about 50,000 cells / cm2, or, more particularly, between about 1,000 cells / cm2 to about 20,000 cells / cm2). If plated at lower densities (e.g. about 300 cells / cm2), the cells can be more easily clonally isolated. For example, after a few days, cells plated at such densities will proliferate into a homogeneous population.

[0062]The high grow rate of the cells of the invention allows quickly and in huge amounts the production of stem cells or conditioned medium. This offers the possibility of (i) carrying out a great number of experiments in order to analyze the biology and uses of these stem cells (this is complicated with the mesenchymal stem cells currently known in the state of the art which show a low grow rate); (ii) a fast tissue regeneration since a high number of stem cell can be obtained in a short period of time; and (iii) quickly obtaining a huge amount of stem cell for its use in anti-inflammatory, anti-tumoral and anti-infectious therapies.

[0065]This allows ensuring the stem cells functionality for a long period of time, being possible to produce a reproducible and effective medicament along stem cell the passages.

[0088]In addition, due to their capacity to regenerate or stimulate tissue regeneration (regenerative medicine), they can be used in wound healing or other processes associated with tissue destruction. Additionally, due to its capacity to secrete substances to the endocervical mucus (which aid the spermatozoa to pass the uterine cervical canal and to reach the ovum), they can be used in fertilization process. As can be seen from the examples, the cells or the conditioned medium of the invention modulate the characteristics of fresh ejaculate and capacitated spermatozoa, helping in the selection of suitable germ cells for fertilization process.

Problems solved by technology

However, except for MAPC, none of these populations has been, until present, sufficiently characterized at the phenotype level.

Therefore, although the presence of multipotent adult cells has been described in different connective tissues, in the current state of the art, it is not possible to identify and unequivocally distinguish between different multipotent cell types obtained from soft tissue, or to obtain a substantially pure population.

Although bone marrow (BM) has been the main source for the isolation of multipotent MSCs, adipose tissue is another source of this cell but the harvest of BM and adipose tissue is a highly invasive procedure.

The process of obtaining bone marrow is painful and the yield is very low, a substantial increase in the number of cells being necessary by ex vivo expansion, to obtain clinically relevant amount.

This step increases cost and makes the procedure time consuming, as well as increases the risk of contamination and loss of material.

Therefore, it is not possible to obtain myometrial tissue by superficial cytological sampling from uterine cervix.

The disadvantage of these stem cells is their contribution to tumor growth.

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