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Stem cell compositions and methods of producing stem cells for therapeutic applications

A stem cell and composition technology, applied in the field of stem cell populations, can solve the problems of reduced cardioprotective effect, limited use, cell aging, etc.

Inactive Publication Date: 2017-08-01
AVITA INT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extensive subcultivation impairs cell function, leading to cellular senescence associated with growth arrest and apoptosis
On the other hand, there are data suggesting that prolonged culture can lead to spontaneous transformation with tumorigenic potential
Furthermore, it was shown that specific properties of MSCs are lost during culture
MSCs were less cardioprotective at passage 5 and above than at passage 10 compared to cells below this passage
Therefore, all methods existing in the prior art should be a compromise between the yield and quality of expanded cells, which limits their use in cell therapy

Method used

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  • Stem cell compositions and methods of producing stem cells for therapeutic applications
  • Stem cell compositions and methods of producing stem cells for therapeutic applications
  • Stem cell compositions and methods of producing stem cells for therapeutic applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1: Vitality of dental pulp

[0081] Maher Al-Atari Abou-Asi et al. (2011) claim that the origin of pluripotent cells extracted from dental pulp (DP) according to the publication of Kerkis et al., 2006 cannot be dental pulp because patients aged 5 to 7 years do not have dental Instead of the pulp, the so-called tooth germ is an aggregate of undifferentiated cells that will form the future tooth with different parts: enamel, pulp, gum, cementum, bone, blood vessels and nerves. However, Cordeiro et al. (2008) found that stem cells from human exfoliated primary teeth generated DP-like tissues in vivo. The authors transplanted SHEDs grown in biodegradable scaffolds prepared within human tooth slices into immunodeficient mice. They observed tissue formation from these cells, demonstrating a structure and cellularity very similar to physiological DP. Ultrastructural analysis of dentin sialoproteins by transmission electron microscopy and immunohistochemistry demonst...

Embodiment 2

[0089] Example 2: Stem cells from dental pulp explants

[0090] Different examples of dental tissue described in detail in a recent review of stem cells derived from dental origin (Deepa Ponnaiyan 2014) describe different characterizations of biomarkers in stem cells derived from dental tissue, such as dental pulp stem cells, periodontal ligament stem cells, stem cells from human Stem cells from exfoliated deciduous teeth, dental follicle progenitor stem cells from the apical papilla, oral periosteum stem cells and most recently from the gingival connective tissue. Strikingly, most markers were similar between different dental stem cells, but the expression of Notch and CD29 markers was different between stem cells from various dental origins. The in vitro and in vivo marker expression of cells obtained by our disclosed method was identical.

Embodiment 3

[0091] Example 3: Human IDPSCs from Teratomas Harvested Repeatedly at Early Passages

[0092] Teratoma formation is an important tool for determining the pluripotency of cells such as embryonic stem cells (ES cells) or induced pluripotent stem cells (iPS cells). Our studies used a consistent protocol similar to that recently published by Gropp et al. (2012) for assessing the teratoma-forming capacity of cells. When 10 6 cells (Gropp et al. used 10 5 cells) of mouse ES cells and human iPS cells subcutaneously co-transplanted into immunodeficient mice with artificial basement membrane (Matrigel), our method was shown to be highly reproducible and efficient. In 100% of cases, we observed teratoma formation in a large number of animals, even at follow-up examinations up to 6 months after transplantation. We used this method for biosafety analysis of other adult mesenchymal stem cells (MSCs), such as those derived from the pulp of primary teeth, umbilical cord, adipose tissue, e...

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Abstract

The present method relates to methods of expanding or increasing stem cell production obtained from donor samples. The methods preferably including the steps of harvesting cells from minimally manipulated tissue using multiply harvesting cycles to increase the number of obtained stem cells.

Description

Background technique [0001] Stem cells are an attractive source of cells for therapeutic applications, medical research, drug testing, and more. Unfortunately, the use of cells as allografts is problematic. Expansion of stem cells to large numbers is a prerequisite for cell therapy, but adult MSCs have limited proliferative capacity. It is common knowledge in the art that the main method used for MSCs isolated from bone marrow (BM) is their ability to adhere to plastic when whole bone marrow is placed in a plastic Petri dish, and after 4 hours , to wash out non-adherent cells. The protocol may also include density centrifugation of BM in a high density solution (eg Ficoll, Percoll) with low viscosity and low osmolarity to obtain a mononuclear fraction of BM containing MSCs. Alternatively, MSCs from solid tissue such as bone can be isolated by placing a small piece of bone in a flask and growing the cells through a bone explant. Alternatively, a collagenase digestion proced...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/0775A61K35/28A61K35/54
CPCA61K35/28A61K35/54C12N5/0605C12N5/0664C12N5/0668A61P29/00A61P37/02C12N2500/02A61K35/12A61K35/50A61K35/51C12N2500/32C12N2500/34C12N2500/38
Inventor I·柯奇斯S·格洛兹曼
Owner AVITA INT
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