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1. (WO2019045649) METHODS FOR ENRICHING MESENCHYMAL STEM CELLS
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CLAIMS

1. A method of enriching non-senescent mesenchymal stem cell (MSCs), the method comprising: contacting a sample comprising MSCs that has been subjected to an in vitro expansion process for MSCs, or a sample comprising MSCs that has not been subjected to an in vitro expansion process for MSCs, or clinical sample comprising MSCs, with a probe that binds to a glycan expressed on the cell surface of MSCs to allow binding of the probe to the glycan; separating non-senescent MSCs bound to the probe from senescent MSCs;

removing the probe from separated non-senescent MSCs; and

- collecting non-senescent MSCs;

wherein separation of senescent MSCs from non-senescent MSC is based on a difference in expression of glycan at the cell surface of MSCs, wherein non-senescent MSCs express more of the glycan than senescent MSCs.

2. The method according to claim 1 , wherein the probe is lectin.

3. The method of claim 2, wherein the lectin is an agglutinin.

4. The method of any one of claims 2 to 3, wherein the lectin is selected from the group consisting of peanut agglutinin (PNA), jacalin (AIL), ricinus communis agglutinin (RCA; Ricin), hairy vetch lectin (VVL), agaricus bisporus lectin, abrus precatorius agglutinin, agropyrum repens lectin, amaranthin, amaranthus leucocarpus lectin, frutalin (derived from the same plant as jacalin), bauhinia purpurea lectin, codium fragile agglutinin, gleheda, lactarius deliciosus lectin, lactarius deterrimus lectin, laelia autumnalis lectin, maclura pomifera agglutinin, bitter gourd seed lectin, morniga-G, sophora japonica lectin, vateirea macrocarpa lectin, vicia graminea lectin, mistletoe lectin I, and combinations thereof.

5. The method of claim 4, wherein the lectin is peanut agglutinin (PNA).

6. The method of any one of the preceding claims, wherein the glycan is an N-linked or an O-linked glycan.

7. The method of claim 6, wherein the glycan is an O-linked glycan.

8. The method of any one of the preceding claims, wherein the glycan is linked via an a-glycosidic bond or a β-glycosidic bond.

9. The method of claim 8, wherein the glycan is linked via a β-glycosidic bond.

10. The method of any one of the preceding claims, wherein the glycan is a 1,3-linked glycan or a

1.4- linked glycan.

11. The method of claim 10, wherein the glycan is a 1,4-linked glycan.

12. The method of any one of claims 8 to 11, wherein the glycan is a i,3-linked glycan or a β1,4- linked glycan.

13. The method of claim 12, wherein the glycan is a i,3-linked glycan.

14. The method of any one of the preceding claims, wherein the glycan is selected from the group consisting of Gai i-4GalNAc i-R (N-(2,4-dihydroxy-6(hydroxymethyl)-5-((3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-3-yl)acetamide), Gal- β 1 ,3- GalNAc (N-(2,5-dihydroxy-6-(hydroxymethyl)-4-((3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-3-yl)acetamide), Gai i- 3GalNAcal-Ser/Thr (0-(3-acetamido-5-hydroxy-6-(hydroxymethyl)-4-((3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)serine/threonine), (Sia)Gai i-3GalNAcal-Ser (5-acetamido-2-((2-((3-acetamido-2-(2-amino-2-carboxyethoxy)-5- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)oxy)-3,5-dihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-4-yl)oxy)-4-hydroxy-6-(l,2,2-trihydroxyethyl)tetrahydro- 2H-pyran-2-carboxylic acid), (Sia)Gai i-3GaiNAcal-Thr, GalNAca-Ser/Thr (0-(3-acetamido- 4.5- dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)serine/threonine) and combinations thereof.

15. The method of claim 14, wherein the glycan is Gal- i,3-GalNAc.

16. The method of any one of the preceding claims, wherein non-senescent MSCs express at least 10%, at least 25%, at least 50%, at least 100%, at least 200% or at least 300% more, or more of the glycan than senescent MSCs.

17. The method of claim 16, wherein non-senescent MSCs express between 10% to 50%, between 20% to 60%, between 30% to 70%, between 40% to 80%, between 50% to 100%, between 90% to 150%, or between 100% to 200% more of the glycan than senescent MSCs.

18. The method according to any one of the preceding claims, wherein the cells are to be used in treatment.

19. A method for performing quality control for non-senescent mesenchymal stem cell (MSCs), the method comprising:

contacting a sample comprising MSCs with a probe that binds to a glycan expressed on the cell surface of MSCs,

determining the amount of non-senescent MSCs present in the sample;

determining the amount of senescent MSCs in the same sample;

wherein the determination of senescent MSCs from non-senescent MSC is based on a difference in expression of glycan at the cell surface of MSCs, wherein non-senescent MSCs express more of the glycan than senescent MSCs; wherein a high amount of non -senescent cells present in the sample is indicative of a sample of acceptable quality; wherein a low amount of non-senescent cells present in the sample is indicative of a sample of inferior quality.

20. The method of claim 19, wherein the sample quality is determined by quantifying the percentage of non-senescent MSCs in the sample.

21. The method of any one of claims 19 to 20, wherein the sample of acceptable quality is considered to have a high percentage of non-senescent MSCs.

22. The method of claim 20, wherein the percentage of non-senescent MSCs is selected from the group consisting of from about 75% to about 100%, from about 89% to about 100%, from about 96% to about 100%, at least about 80%, at least about 85%, at least about 87%, at least about 90%, at least about 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least about 99.9% of non-senescent MSCs to senescent MSCs.

23. The method of claim 22, wherein the percentage is at least 90% non-senescent MSCs in a sample.