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Last Updated: May 4, 2024

Claims for Patent: 7,754,479

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Summary for Patent: 7,754,479

Title:	Method for culturing avian spermatogonial stem cells and avian spermatogonial stem cells prepared thereby
Abstract:	The present invention relates to a method for long-term culturing of avian spermatogonial stem cells, which comprises the steps of: (a) preparing an avian testis; (b) isolating a population of testicular cells from said avian testis; and (c) culturing said avian spermatogonial stem cells in said population of testicular cells on a feeder cell layer in a medium containing a cell growth factor, a population of avian spermatogonial stem cells and a method for producing transgenic ayes.
Inventor(s):	Han; Jae Yong (Seoul, KR), Hong; Yeong Ho (Suwon-si, KR), Lim; Jeong Mook (Seoul, KR), Lee; Young Mok (Suwon-si, KR), Lee; Mak Soon (Suwon-si, KR), Jung; Jin Gyung (Seoul, KR)
Assignee:	Seoul National University Industry Foundation (Seoul, KR) Avicore Biotechnology Institute Inc. (Gunpo, KR)
Application Number:	10/569,847
Patent Claims:	1. A method for a long-term culture for more than 3 months of avian spermatogonial stem cells, which comprises the steps of: (a) preparing an avian testis from an avian aged 2-70 weeks; (b) isolating a population of testicular cells from said avian testis; (c) culturing said population of testicular cells for about 5 to 10 days on plates in a medium containing a cell growth factor to form a colony of spermatogonial stem cells; and (d) taking a colony of spermatogonial stem cells and culturing said avian spermatogonial stem cells for more than about 80 to 85 days on a feeder cell layer in a medium containing a cell growth factor; wherein said step (b) is carried out by treating said avian testis with a mixture of collagenase and trypsin; said medium in steps (c) and (d) includes FBS (fetal bovine serum), avian serum, non-essential amino acids, Hepes buffer, and .beta.-mercaptoethanol; and said feeder cell is avian gonadal stroma cell or testicular stroma cell. 2. The method according to claim 1, wherein said cell growth factor is a growth factor selected from the group consisting of fibroblast growth factor, insulin-like growth factor-1, stem cell factor, glial-derived neurotrophic factor and their combination. 3. The method according to claim 1, wherein said medium further comprises a differentiation inhibitory factor. 4. The method according to claim 3, wherein said differentiation inhibitory factor is leukemia inhibitory factor. 5. The method according to claim 1, wherein said medium comprises a supplement containing a mixture of fibroblast growth factor, insulin-like growth factor-i and leukemia inhibitory factor. 6. The method according to claim 1, wherein said medium further comprises a serum and an antioxidant. 7. The method according to claim 1, wherein said culturing is carried out at about 37.degree. C. 8. The method according to claim 1, wherein said avian species is a chicken, a quail, a turkey, a duck, a goose, a pheasant or a pigeon. 9. The method according to claim 1, wherein after step (c) said process further comprises the step of identifying the avian spermatogonial stem cells. 10. The method according to claim 9, wherein said identification is carried out by (i) PAS (Periodic Acid Shift's) staining, (ii) STA (Solanum tubersum agglutinin) staining, (iii) a staining with .alpha.6-integrin antibody, (iv) a staining with .beta.1-integrin antibody, (v) a staining with anti-S SEA-1 antibody, (vi) a staining with anti-SSEA-3 antibody, (vii) a staining with anti-S SEA-4 antibody, (viii) DBA (Doliclos biffirus agglutinin) staining or (ix) their combination. 11. The method of claim 1, wherein said avian is aged up to 20 weeks. 12. The method of claim 11, wherein said avian is aged 2-10 weeks. 13. The method of claim 1, wherein said avian is a chicken. 14. The method of claim 12, wherein said avian is a chicken. 15. A method for a long-term culture for more than 3 months of avian spermatogonial stem cells, which comprises the steps of: (a) preparing an avian testis from an avian that is not in an embryonic stage; (b) isolating a population of testicular cells from said avian testis; and (c) culturing said population of testicular cells for about 5 to 10 days on plates in a medium containing a cell growth factor to form a colony of spermatogonial stem cells; and (d) taking a colony of spermatogonial stem cells and culturing said avian spermatogonial stem cells for more than about 80 to 85 days on a feeder cell layer in a medium containing a cell growth factor; wherein said step (b) is carried out by treating said avian testis with a mixture of collagenase and trypsin; said medium in steps (c) and (d) includes FBS (fetal bovine serum), avian serum, non-essential amino acids, Hepes buffer, and .beta.-mercaptoethanol; and said feeder cell is avian gonadal stroma cell or testicular stroma cell.

Details for Patent 7,754,479

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Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965	⤷ Try a Trial	2023-08-08
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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