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Stem cells expressing mesenchymal and neuronal markers, compositions thereof, and methods of preparation thereof

A composition and technology of dental pulp stem cells, applied in the field of neurological diseases

Pending Publication Date: 2019-02-05
AVITA INT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite research on stem cell therapy, there are no data showing that stem cell therapy via intravenous (IV) injection can directly lead to neurogenesis through secretion of BDNF or expression of D2 in the ventricles of neurodegenerative diseases

Method used

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  • Stem cells expressing mesenchymal and neuronal markers, compositions thereof, and methods of preparation thereof
  • Stem cells expressing mesenchymal and neuronal markers, compositions thereof, and methods of preparation thereof
  • Stem cells expressing mesenchymal and neuronal markers, compositions thereof, and methods of preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0176] Example 1: Characterization of Early and Late Harvest IDPSCs and Derivation of Neural and Glial Cells from Early and Late Harvest IDPSCs

[0177] Characterization of Early and Late Harvest IDPSCs

[0178] To characterize hIDPSC properties, early (n=8) and late (n=4) harvests (Table 1) were harvested for flow cytometric analysis of mesenchymal markers CD13, CD105, CD73, CD90, CD44. Using 2×10 5 cells for FACS experiments. Cells were washed twice with PBS (without calcium and magnesium), and the tested antibodies were added for 15 minutes at room temperature. Cells were then washed twice with cold PBS and analyzed by Becton-Dickinson flow cytometry. The fluorescence of PE(FL2), FITC(FL1), APC(FL4) was detected at emission wavelengths of 575nm, 53nm and 600nm, respectively.

[0179] Table 1. List of cells used in FACS experiments

[0180]

[0181] Cells from both early and late harvests expressed high levels of mesenchymal markers. Both populations are negative fo...

Embodiment 2

[0198] Example 2. Expression of CD146 and CD13 in early stage (EP) and late stage (LP) hIDPSCs

[0199] CD146 and CD13 expression was analyzed by flow cytometry in EP-hIDPSCs and LP-hIDPSCs. Figure 7 The results in showed that CD13 was expressed in 52% of EP-hIDPSCs and 95% of LP-hIDPSCs. These results suggest that in vitro DP harvest and hIDPSC passaging yields increased numbers of hIDPSCs expressing CD13 and lacking CD146 expression.

Embodiment 3

[0200] Comparison of embodiment 3.IDPSC and SHED

[0201] MSCs from different sources such as bone marrow and adipose tissue respond differently to different stimuli (Fraser JK et al., 2006). Culture conditions (eg, medium supplemented with human or fetal calf serum (FCS) or serum-free) may also affect the differentiation potential of MSCs even of the same origin (Lindroos et al., 2011; Lizier et al., 2012). Differences in differentiation efficiency likely largely reflect the heterogeneity of the MSC population (ie, the presence of different subpopulations) (Ho et al., 2008; Tormin et al., 2009; Mareddy et al., 2009; Rada et al., 2011). Different isolation and culture protocols used by different populations may explain the predominance of specific MSC subpopulations with different differentiation potential (Ho et al., 2008; Pevsner-Fischer et al., 2011; Rada et al., 2011).

[0202] SHED and IDPSC have different isolation methods and come from different stem cell niches. Ther...

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Abstract

The invention provides a method of producing human immature dental pulp stem cells (hIDPSCs) expressing CD44 and CD13 and lacking expression of CD146. The invention also provides compositions for usein the treatment of a neurological disease or condition selected from the group consisting of Parkinson's disease (PD), multiple sclerosis, 5 amyotrophic lateral sclerosis (ALS), stroke, autoimmune encephalomyelitis, diabetic neuropathy, glaucomatous neuropathy, Alzheimer's disease, Huntington's disease (HD), autism, schizophrenia, stroke, ischemia, a motor disorder, and a convulsive disorder.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Patent Application No. 15 / 065,259 (published as US2016 / 0184366), filed March 9, 2016, which is hereby incorporated by reference in its entirety. technical field [0003] The present application relates to methods of producing stem cells, stem cells and compositions comprising stem cells suitable for the treatment of several diseases, especially neurological diseases, suitable for systemic administration. Background technique [0004] Although the genes responsible for neurodegenerative diseases and their proteins have been identified, the mechanisms involved in the pathogenesis of these diseases remain unknown, hindering the development of effective therapeutic interventions. What is known is that despite its ubiquity, for example, mutant forms of huntingtin cause neurodegeneration and selective loss of medium spiny neurons, which occur preferentially in the striatum and deeper la...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/32A61P25/00A61P25/28
CPCA61K35/32C12N5/0664A61P21/02A61P25/00A61P25/08A61P25/14A61P25/16A61P25/18A61P25/28A61P27/02A61P27/06A61P35/00A61P37/06A61P9/00A61P9/10A61K9/0019A61K38/177C12N5/00
Inventor I·柯奇斯C·瓦尔韦德文塞斯劳
Owner AVITA INT
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