Claims for Patent: 9,241,959

Summary for Patent: 9,241,959

Title:	Kits and methods for processing stem cells from bone marrow or umbilical cord blood
Abstract:	Provided herein are kits for processing stem cells from bone marrow or umbilical cord blood, comprising: a) a precipitation reagent comprising an aqueous solution of 0.1-30% hydroxyethyl starch or 0.1-30% methyl cellulose, and 0.1-20% of cerebroprotein hydrolysate; and b) a separation reagent comprising an aqueous solution comprising Ficoll and diatrizoate and having a density of 1.0-1.2 g/ml, and methods of use. Further provided herein is a collection of stem cells obtained by the kits and methods disclosed herein comprising stem cells from bone marrow or umbilical cord blood, and uses thereof.
Inventor(s):	Tang; Mingqi (Beijing, CN)
Assignee:	Tang; Mingqi (Beijing, CN)
Application Number:	14/203,152
Patent Claims:	1. A method for separating stem cells using a kit for processing stem cells from bone marrow or umbilical cord blood comprising a precipitation reagent comprising an aqueous solution of 0.1-30% hydroxyethyl starch or 0.1-30% methyl cellulose, and 0.1-20% of cerebroprotein hydrolysate; and a separation reagent comprising an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of 1.0-1.2 g/ml, which method comprises: a) providing a bone marrow or umbilical cord blood sample; b) adding the precipitation reagent to the bone marrow or umbilical cord blood sample, mixing and letting the solution settle, followed by removal and centrifugation of the upper layer to obtain a concentrated sample; and c) adding the concentrated sample from step b) on top of the separation reagent and performing density gradient centrifugation to obtain a layer comprising the stem cells. 2. The method of claim 1, wherein step a) further comprises adding an anticoagulant to the bone marrow or umbilical cord blood. 3. The method of claim 1, wherein step a) further comprises adding an equal volume of saline (NaCl injection) to the bone marrow or umbilical cord blood sample to obtain a diluted sample. 4. The method of claim 3, wherein the ratio between the diluted sample and the precipitation reagent is 2:1 by volume. 5. The method of claim 1, wherein step a) is performed in a sterile environment. 6. The method of claim 1, wherein the bone marrow is obtained using a bone marrow needle from the posterior superior iliac spine with anesthesia. 7. The method of claim 1, wherein the concentrated sample from step b) is diluted with saline. 8. The method of claim 1, wherein the bone marrow or umbilical cord blood sample is freshly collected from a mammal. 9. The method of claim 8, wherein the mammal is a human being. 10. The method of claim 1, wherein the bone marrow or umbilical cord blood sample has not been cultivated ex vivo. 11. The method of claim 1, wherein the precipitation reagent comprises 0.3-15% of cerebroprotein hydrolysate. 12. The method of claim 1, wherein the precipitation reagent comprises 0.8-6% of cerebroprotein hydrolysate. 13. The method of claim 1, wherein the precipitation reagent comprises about 1.5-4.5% of cerebroprotein hydrolysate. 14. The method of claim 1, wherein the precipitation reagent comprises about 3% of cerebroprotein hydrolysate. 15. The method of claim 1, wherein the precipitation reagent comprises 0.5-18% aqueous solution of hydroxyethyl starch or 0.2-8% aqueous solution of methyl cellulose. 16. The method of claim 1, wherein the precipitation reagent comprises about 1.5-12% aqueous solution of hydroxyethyl starch or about 0.3-5% aqueous solution of methyl cellulose. 17. The method of claim 1, wherein the precipitation reagent comprises about 6% aqueous solution of hydroxyethyl starch or about 0.5% aqueous solution of methyl cellulose. 18. The method of claim 1, wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.01-1.088 g/ml. 19. The method of claim 1, wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.035-1.08 g/ml. 20. The method of claim 1, wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.075 g/ml. 21. The method of claim 1, wherein the precipitation reagent comprises 0.5-18% aqueous solution of hydroxyethyl starch or 0.2-8% aqueous solution of methyl cellulose, and 0.8-6% of cerebroprotein hydrolysate, and wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.01-1.088 g/ml. 22. The method of claim 1, wherein the precipitation reagent comprises about 1.5-12% aqueous solution of hydroxyethyl starch or about 0.3-5% aqueous solution of methyl cellulose, and about 1.5-4.5% of cerebroprotein hydrolysate, and wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.035-1.08 g/ml. 23. The method of claim 1, wherein the precipitation reagent comprises about 6% aqueous solution of hydroxyethyl starch or about 0.5% aqueous solution of methyl cellulose, and about 3% of cerebroprotein hydrolysate, and wherein the separation reagent comprises an aqueous solution comprising Ficoll polysaccharide and diatrizoate and having a density of about 1.075 g/ml. 24. The method of claim 1, wherein the aqueous solution uses sterile water suitable for injection into a subject. 25. The method of claim 24, wherein the subject is a human being. 26. The method of claim 1, wherein the precipitation reagent and the separation reagent are sterilized under ultrafiltration conditions.

Details for Patent 9,241,959

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	ZOSTAVAX	zoster vaccine live	For Injection	125123	05/25/2006		2033-03-13