Tissue culture medium

• US 4,533,634 A
• Filed: 01/26/1983
• Issued: 08/06/1985
• Est. Priority Date: 01/26/1983
• Status: Expired due to Term

• Current Assignee: Sigma-Aldrich Company (Sigma-Aldrich Corporation)
• Original Assignee: AMFM Incorporated
• Inventors: Maldonado, Reynaldo L., Rosanoff, Kenneth A.
• Primary Examiner(s): Wiseman, Thomas G.
• Assistant Examiner(s): Huleatt, Jayme A.
• Application Number:
  US06/461,016
• Time in Patent Office: 923 Days
• Field of Search: 435/240, 435/241, 435/244, 435/243, 260/112 B, 424/101
• US Class Current: 435/392
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 5/00 Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00
• CPC Class Codes: Y10S 530/808 Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
• CPC Class Codes: Y10S 530/83 Plasma; serum

Abstract

A growth factor containing serum is disclosed which is derived from natural bovine serum is disclosed which is by precipitating a first solids fraction from natural bovine serum by adding ammonium sulfate to about 25% saturation to give a first solids fraction and a first supernatant; separating the first solids fraction from the first supernatant; precipitating a second solids fractions from the first supernatant by adding ammonium sulfate to about 40% saturation to give a second solids fraction and a second supernatant; separating the second solids fraction from the second supernatant; combining the first solids fraction and the second supernatant; and desalinating the first solids fraction and the second supernatant, either before or after combination thereof.

Claims

1. A process for producing a growth factor containing serum having a γ

-globulin content of between about 0.1-1.0 g/dl, and of 50-100 mg/dl of cholesterol comprising the steps of;

(a) adding ammonium sulfate to natural bovine serum to 15-30% saturation to give a first solids fraction and a first supernatant;

(b) separating said first solids fraction from said first supernatant;

(c) adding ammonium sulfate to said first supernatant to 35-45% saturation to give a second solids fraction and a second supernatant;

(d) separating said second solids fraction from said second supernatant;

(e) combining said first solids fraction and said second supernatant; and

(f) desalinating said first solids fraction and said second supernatant, either before or after said combining.

2. The process of claim 1, wherein said first solids fraction is obtained by adding ammonium sulfate to 20-28% saturation.

3. The process of claim 1, wherein said first solids fraction is obtained by adding ammonium sulfate to 25% saturation.

4. The process of claim 1, wherein said second solids fraction is obtained by adding ammonium sulfate to 38-42% saturation.

5. The process Of claim 1, wherein said second solids fraction is obtained by adding ammonium sulfate to 40% saturation.

6. The process of claim 1 which further comprises contacting said serum with charcoal.

7. The process of claim 1 which further comprises sterilizing said serum.

8. The process of claim 1 which further comprises heat inactivating said serum.

9. A method of enhancing the cell growth promoting ability of natural bovine serum which comprises treating said serum by the process of claim 1.