Human uterine cervical stem cell population and uses thereof

a human uterine cervical and stem cell technology, applied in the field of human uterine cervical stem cell population, can solve the problems of inability to identify and unequivocally distinguish between different multipotent cell types obtained from soft tissue, inability to obtain a substantially pure population, and high invasive procedure for harvesting bm and adipose tissu

Inactive Publication Date: 2016-01-07
GISTEM RESERCH SL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention relates to a new source of stem cells, called uterine cervical stem cells (UCESC), which can be obtained from the tissue of the uterine cervix. These cells show a higher anti-inflammatory, anti-tumor, antimicrobial, and growth rate compared to other stem cells. Additionally, they can be obtained non-invasively and painlessly from the same organ during a routine gynaecological examination. The cells can be expanded in culture and differentiated into different tissue types, making them useful for various regenerative medicine applications such as tissue destruction, inflammation, tumor treatment, and fertilization process. The high grow rate of the cells enables quick and in huge amounts the production of stem cells, which can be used for experimentation, tissue regeneration, and medicament production. The stem cells can also be modulated to improve their characteristics for fertilization.

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.

Method used

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  • Human uterine cervical stem cell population and uses thereof
  • Human uterine cervical stem cell population and uses thereof
  • Human uterine cervical stem cell population and uses thereof

Examples

Experimental program
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example 1

Isolation and Characterization of Human Uterine Cervical Stem Cells

I—Material and Methods

Isolation and Growth of Human Uterine Cervical Stem Cells

[0137]Human uterine cervical stem cells (hUCESCs) were obtained from an exfoliation of the uterine cervix during routine gynaecological examination. Briefly, cytological sample was enzymatically disaggregated with trypsin, collagenase or other enzyme which can disaggregate the cervical mucus. Then, the sample was centrifuged 5 minutes at 400 g and the pellet was collected and seeded in a culture plate. The well can be previously coated with 1% gelatin or fibronectin or other substrate to allow the adherence. Sample was culture in Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM-F12), glutamine, with or without antibiotics, with serum, epidermal growth factor (EGF), hydrocortisone, insulin, non-essential amino acids, sodium pyruvate. The subculture of cells was carried out with trypsin or accutase or other proteolytic and / or co...

example 2

Inflammation Related Experiments

I—Material and Methods

Immunogenicity Assay

[0147]The one-way mixed lymphocyte reaction (MLR) assay was used to determine the immunogenicity of human uterine cervical stem cells (hUCESCs). The MLR was performed in 96-well microtiter plates using RPMI 1640 medium without FBS. Peripheral blood mononuclear cells (PBMC) derived from two different donors were plated at 2×105 cells per donor per well. Different donors were used to maximize the chance that at least one of PBMC was a major mismatch to the hUCESCs test cells. Stimulator cells used in the assay included autologous PBMC (baseline response), allogeneic PBMC (positive-control response), and hUCESCs cell population. Stimulator cells were mitomycin C treated prior to being added to the culture wells (2×104 cells per well, 10% stimulators cells). Additional controls cultures consisted of PBMC plated in medium alone (no stimulator cells), concanavalin A (ConA) stimulated PBMC and of hUCESCs mitomycin C ...

example 3

Cancer Related Experiments

I.—Material and Methods

Cell Cultures

[0154]MCF-7 and MDA-MB-231 cells (human breast adenocarcinoma cell lines) were obtained from the European Collection of Cell Cultures (Salisbury, Wilts., UK) and HT29 (colorectal adenocarcinoma cell line) and AGS (gastric adenocarcinoma cell line) were obtained from the American Type Culture Collection (ATCC, Manassas, Va., USA). These cell lines were grown in 90-mm Petri dishes in DMEM supplemented with 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin in an air-CO2 (95:5) atmosphere at 37° C. Confluent cells were washed twice with phosphate-buffered saline and harvested by a brief incubation with trypsin-EDTA solution (Sigma-Aldrich, St. Louis, Mo., USA) in PBS. Human cervical uterine stem cells (hUCESCs, obtained as above described), primary cultures from human breast tumours, and human adipose-derived stem cells (ASCs, StemPro®, Invitrogen), were grown in 90-mm Petri dishes in DMEM-F12 (1:1) supplemented with 1...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/48C12N5/074
CPCA61K35/48C12N2502/03C12N2509/00C12N5/0607C12N5/0661C12N5/0668C12N5/0682A61P15/00A61P17/00A61P29/00A61P31/04A61P35/00A61P35/04A61P37/06A61P43/00A61P7/00
Inventor VIZOSO PINEIRO, FRANCISCO JOSEPEREZ FERN NDEZ, ROMANEIRO D AZ, NOEMI
Owner GISTEM RESERCH SL
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