Claims for Patent: 10,273,457

Summary for Patent: 10,273,457

Title:	Method of obtaining a population of cells
Abstract:	A method of obtaining stromal progenitor cells (SPC) from subcutaneous adipose tissue by incubation of a very small volume of the subcutaneous adipose tissue in an enzyme solution produces SPC that are usable in medical applications based on autologous SPC even on individuals having a body mass index lower than 18.5.
Inventor(s):	Dominici; Massimo (Ferrara, IT), Cafarelli; Luigi (Modena, IT), Veronesi; Elena (Finale Emilia, IT), Piccinno; Maria Serena (Modena, IT), Paolucci; Paolo (Bologna, IT), De Santis; Giorgio (Modena, IT), Conte; Pierfranco (Pisa, IT)
Assignee:
Application Number:	14/309,086
Patent Claims:	1. A method of obtaining a population of stromal progenitor cells comprising the steps of: collecting a first quantity q.sub.1 of subcutaneous adipose tissue containing white adipose cells from a living being; digesting the first quantity q.sub.1 with a second quantity q.sub.2 of enzymatic solution comprising collagenase to obtain a population of free cells; seeding the population of free cells into a culture medium; and obtaining stromal progenitors cells from the seeded cells, wherein q.sub.1 is between 0.004 ml and 0.036 ml and q.sub.2 is between 0.9 ml and 1.1 ml, and wherein q.sub.1 and q.sub.2 can be proportionally scaled up or down. 2. The method according to claim 1, wherein said enzymatic solution comprises: Dulbecco's Modified Eagle Medium (DMEM); penicillin (1 U/ml) & streptomycin (1 mg/ml); sodium pyruvate (1 mM); and non-essential amino acids comprising: L-alanine (0.89 mg/dl); L-asparagine H2O (1.5 mg/dl); L-aspartic Acid (1.33 mg/dl); L-glutamic Acid (1.47 mg/dl); glycine (0.75 mg/dl); L-proline (1.15 mg/dl); and L-serine (1.05 mg/dl). 3. The method according to claim 1, wherein said enzymatic solution comprises: class I and II collagenase mixture; or class I and II collagenase mixture with an enzymatic concentration between 1.74 and 1.78 U/ml of said enzymatic solution. 4. The method according to claim 1, wherein said enzymatic solution comprises: clostripain; and trypsin. 5. The method according to claim 1, wherein the step of digesting comprises joining said first and second quantities q.sub.1 and q.sub.2together, and, between the steps of collecting and seeding, shaking said first and second quantities q.sub.1 and q.sub.2 with a shaking device for a time between 20 minutes and 120 minutes at a predetermined temperature. 6. The method according to claim 5, wherein said predetermined temperature is between 36.degree. C. and 38.degree. C. 7. The method according to claim 5, wherein the step of shaking comprises swinging said shaking device in a first swinging direction with a first number of swings per minute and in a second swinging direction different from said first swinging direction and with a second number of swings per minute. 8. The method according to claim 7, wherein said shaking device comprises a tubular container. 9. The method according to claim 8, wherein said shaking device is adapted to move said tubular container in a direction of a longitudinal axis of said tubular container. 10. The method according to claim 7, wherein said first number is between 70 and 80 swings per minute and said second number is between 6 and 100 swings per minute. 11. The method according to claim 1, wherein said stromal progenitor cells comprise mesenchymal stromal cells and stromal pericytes. 12. The method according to claim 1, wherein seeding the population of free cells into a culture medium further comprises the step of expanding the population of free cells in the culture medium, wherein the culture medium comprises a synthetic basal medium with an antibiotic added therein.

Details for Patent 10,273,457

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2030-04-14