PURIFIED MESENCHYMAL STEM CELL COMPOSITIONS AND METHODS OF PURIFYING MESENCHYMAL STEM CELL COMPOSITIONS
One or more purified mesenchymal stem cell pharmaceutical compositions and methods of manufacture utilizing centrifugal filtration are disclosed. Threshold limits for intravenous administration of mesenchymal stem cell pharmaceutical compositions comprising residual animal products are also disclosed.
1 . A method of preparing a pharmaceutically acceptable composition comprising a plurality of mesenchymal stem cells and mesenchymal stem cell aggregates comprising the steps of:
(i) obtaining a mesenchymal stem cell suspension comprising a plurality of mesenchymal stem cells and mesenchymal stem cell aggregates;
(ii) contacting the suspension with a wash solution to create a mixed suspension;
(iii) agitating the mixed suspension with a centrifugal filtration device until the D 90 of the mesenchymal stem cell aggregates in the mesenchymal stem cell suspension is less than about 150 μm;
(iv) recovering the pharmaceutically acceptable composition.
2 . The method of claim 1 , wherein the mesenchymal stem cell aggregates exhibit the D 90 of less than about 100 μm.
3 . The method of claim 2 , wherein the mesenchymal stem cell aggregates exhibit the D 90 of less than about 50 μm.
4 . The method of claim 1 , wherein the mesenchymal stem cell aggregates exhibit a D 90 between about 18 μm and about 30 μm.
5 . The method of claim 4 , wherein the mesenchymal stern cell aggregates exhibit a D 90 between about 18 μm and about 25 μm.
6 . The method of claim 5 , wherein the mesenchymal stem cell aggregates exhibit a D 90 between about 20 μm and about 25 μm.
7 . The method of claim 1 , further comprising DMSO.
8 . The method of claim 7 , further comprising about 10% DMSO.
9 . The method of claim 7 , further comprising about 3.8% DMSO.
10 . The method of claim 1 wherein the viability of the cells is greater than about 70%.
11 - 24 . (canceled)
25 . The method of claim 1 , wherein the mesenchymal stem cell suspension is cultured in media comprising bovine serum albumin.
26 . The method of claim 1 , wherein the pharmaceutically acceptable composition comprises less than about 55 μg/mL residual bovine serum albumin.
27 . The method of claim 26 , wherein the pharmaceutically acceptable composition comprises less than about 25 μg/mL bovine serum albumin.
28 . The method of claim 27 , wherein the pharmaceutically acceptable composition comprises between about 8 μg/mL and about 12 μg/mL bovine serum albumin.
29 . A method of preventing pulmonary embolism in a subject, wherein pulmonary embolism is caused by aggregation of mesenchymal stem cells, comprising administering a pharmaceutically acceptable composition comprising a plurality of mesenchymal stem cells and mesenchymal stem cell aggregates, wherein the aggregates comprise a D 90 of less than about 150 μm.
30 . The method of claim 29 , wherein the mesenchymal stem cell aggregates exhibit a D 90 of less than about 100 μm.
31 . The method of claim 30 , wherein the mesenchymal stem cell aggregates exhibit a D 90 of less than about 50 μm.
32 . The method of claim 29 , wherein the mesenchymal stem cell aggregates exhibit a D 90 between about 18 μm and about 30 μm.
33 . The method of claim 32 , wherein the mesenchymal stem cell aggregates exhibit a D 90 between about 18 μm and about 25 μm.
34 . The method of claim 33 , wherein the mesenchymal stem cell aggregates exhibit a D 90 between about 20 μm and about 25 μm.
35 . The method of claim 29 , wherein the pharmaceutically acceptable composition comprises less than about 55 μg/mL bovine serum albumin.
36 . The method of claim 35 , wherein the pharmaceutically acceptable composition comprises less than about 25 μg/mL bovine serum albumin.
37 . The method of claim 36 , wherein the pharmaceutically acceptable composition comprises between about 8 μg/mL and about 12 μg/mL bovine serum albumin.
38 . The method of claim 29 , wherein viability of the cells is greater than about 70%.
39 . The method of claim 29 , wherein the subject is treated in response to an immune response, vasoconstriction, or cardiac shock.