Title:
HUMAN EMBRYONIC KIDNEY (HEK) 293 SUBCLONE CELL STRAIN
Document Type and Number:
WIPO Patent Application WO/2004/111239
Kind Code:
A1
Abstract:
The present invention disclosed a HEK 293 subclone cell strain for producing recombinant adenovirus preparation, the said cell strain, which comprises human glyceraldehydes-3-phosphate dehydrogenase (GAPD), is made of ATCC 293 cell via subcloning method. The producing method includes the following steps: a. proliferating the ATCC 293 cell (P32) to P33, and then using FACS flow cytometer to subclone the P33 suspension; b. using two 96-well plates, wherein placing a cell in 0.2ml medium of every cell, and culturing at 3638C, 3-6 % C02 for 6- 10days; c. selecting the clones with the cell uniform, growth well and not less than 60% confluent culture; d. repeating the above steps 4times, and finally selecting a cell strain(P37) of subclone, continuing to amplify and obtain the proto-cell library(P38), and freezing under - 80'C; e. preparing the parental cell library and working cell library based on proto-cell library. The present invention makes it having the advantages of increasing the function of the cells adherence growth and inducing the multiplication time of cell growth substantially.
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Inventors:
ZHANG XIAOZHI (CN)
PENG ZHAOHUI (CN)
PENG ZHAOHUI (CN)
Application Number:
PCT/CN2004/000457
Publication Date:
December 23, 2004
Filing Date:
May 09, 2004
Export Citation:
Assignee:
ZHANG XIAOZHI (CN)
PENG ZHAOHUI (CN)
PENG ZHAOHUI (CN)
International Classes:
C12N5/071; C12N5/073; C12N15/50; (IPC1-7): C12N15/50; C12N5/00; C12N9/04
Other References:
DATABASE GENBANK [online] 8 September 2000 (2000-09-08), YE Z. ET AL., XP002982880, accession no. NCBI Database accession no. (AF261085)
GUAN Y. ET AL.: "Study in the Ca2+ dependent K+ current and Ca2+ entry induced by activation of alpha1B adrenoceptor subtype in HEK-293-cell", ACAD. J. SUMS, vol. 20, no. 4, 1999, pages 248 - 251, XP008042018
WANG Y. ET AL.: "Funcional analysis of the human T-type calcium channel alpha1H subunit gene in cellular proliferation", ACTA GENETICA SINICA, vol. 29, no. 8, 2002, pages 659 - 665, XP008042020
GUAN Y. ET AL.: "Study in the Ca2+ dependent K+ current and Ca2+ entry induced by activation of alpha1B adrenoceptor subtype in HEK-293-cell", ACAD. J. SUMS, vol. 20, no. 4, 1999, pages 248 - 251, XP008042018
WANG Y. ET AL.: "Funcional analysis of the human T-type calcium channel alpha1H subunit gene in cellular proliferation", ACTA GENETICA SINICA, vol. 29, no. 8, 2002, pages 659 - 665, XP008042020
Attorney, Agent or Firm:
JEEKAI & PARTNERS (Jinyu Tower A12, West Xuan Wu Men Street Beijing 1, CN)
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