Claims for Patent: 5,543,318

Summary for Patent: 5,543,318

Title:	Method of isolation, culture and proliferation of human atrial myocytes
Abstract:	The present invention provides methods for isolating and culturing human myocardiocytes from adult human atrial tissue and proliferating cultures resulting therefrom. In one embodiment, the myocardial cells are substantially dissociated through sequential enzymatic digestion and plated on growth medium selective for myocardial cell growth. In another embodiment, the atrial tissue is subjected to only limited digestion to provide an explant tissue portion which is cultured under conditions favoring cell proliferation. Cells migrating from the explant are isolated and subjected to time-based selective attachment and selective growth techniques which result in a myocardial cell culture of approximately 93% purity.
Inventor(s):	Smith; David A. (Royal Oak, MI), Townsend; Laurace E. (Grosse Pointe Park, MI)
Assignee:
Application Number:	08/531,793
Patent Claims:	1. A method of reproducing human atrial mycocardiocytes in culture comprising the steps of: providing a sample of human tissue comprising adult atrail cells; growing at least a portion of the cells in the samples in culture medium to obtain a cell culture; selecting for myocardinocytes in the culture; and maintaining the selected myocardiocytes in culture for a time sufficient and under culture conditions which allow for cell division of the myocardiocytes. 2. The method of claim 1, further comprising the step of partial dissociation of the cells in the tissue sample, wherein after the step of partial dissociation the sample comprises an explant tissue portion and a diassociated cell portion, and wherein the explant tissue portion comprises the portion grown in culture medium. 3. The method of claim 2, wherein after the step of dissociation the sample comprises substantially dissociated cells and the portion grown in culture medium comprises substantially dissociated cells. 4. The method of claim 3, wherein the step of selecting comprises growth of the substantially dissociated cells in growth medium selective for myocardiocyte growth. 5. The myocardiocyte culture produced by the method of claim 4. 6. The method of claim 2, wherein the explant tissue portion is cultured on a cell attachment matrix under conditions favorable for explant attachment, cell proliferation and migration from the explant tissue portion. 7. The method of claim 6, further comprising the step of isolating cells migrating from the explant tissue portion, wherein the isolated cells include myocardiocytes, and wherein the step of selecting further comprises the steps of placing the isolated cells on a cell attachment matrix for a time period sufficient for isolated myocardiocytes to attach to the matrix and removing unattached cells after the expiration of the time period. 8. The myocardinocyte culture produced by the method of claim 7. 9. The method of claim 7, wherein the step of selecting further comprises growth of the cells selected by attachment in growth medium selective for myocardiocyte growth. 10. The myocardiocyte culture produced by the method of claim 9. 11. The method of claim 1, wherein the step of maintaining the myocardiocytes in culture further comprises the step of growing the selected myocardiocytes to confluence in culture and passing the confluent myocardiocytes. 12. The myocardiocyte culture produced by the method of claim 1. 13. A method of reproducing human atrail myocardinocytes in culture comprising the steps of: providing a sample of human atrail tissue comprising an explant tissue portion further comprising adult cells; growing the explant tissue portion in culture medium to obtain a cell culture; isolating cells migrating from explant tissue portion wherein the cells isolated include myocardiocytes; plating the isolated cells on a cell attachment matrix; selecting for myocardiocytes wherein the step of selecting further comprises the steps of maintaining the isolated cells in the cell attachment matrix for a time period sufficient for at least a portion of myocardiocytes present in the isolated cells to attach to the matrix and removing unattached cells after expiration of the time period; and maintaining the selected myocardiocytes in culture for a time sufficient for cell division to occur. 14. The method of claim 13, further comprising the step of dissociating a portion of the cells of the sample to provide the explant tissue portion and a dissociated cell portion. 15. The method of claim 14, wherein the step of dissociation further comprises partial dissociation of the sample through enzymatic digestion of the tissue sample. 16. The method of claim 13, wherein the time period is approximately 30 minutes. 17. The myocardinocyte culture produced by the method of claim 13. 18. A method of reproduce human atrial myocardiocytes in culture comprising the steps of: providing a sample of adult human atrail tissue; substantially dissociating the cells in the samples; growing the dissociated cells in culture medium to provide a cell culture; selecting for myocardiocytes in at least a portion of the disassociated cells; and growing the selected myocardinocytes in culture for a time sufficient to allow cell division to occur in at least a portion of the myocardiocytes. 19. The method of claim 18, wherein the step of substantial dissociation further comprises sequential digestion by an enzyme for a time period in the range of approximately two to nine hours. 20. The method of claim 19, wherein the enzyme used for digestion is collagenase and the time period of digestion is in the range of approximately six to seven hours. 21. The method of claim 18, wherein the step of selecting comprises culturing the dissociated cells in growth medium selective for myocardiocyte growth. 22. The myocardiocyte culture produced by the method of claim 21. 23. The myocardinocyte culture produced by the method of claim 18.

Details for Patent 5,543,318

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2011-06-12