Synthetic tissue controls and synthetic tissue microarray controls

• US 10,329,623 B2
• Filed: 10/12/2015
• Issued: 06/25/2019
• Est. Priority Date: 08/28/2015
• Status: Active Grant

• Current Assignee: Slmp, LLC (SLMP Holding LLC), Statlab Medical Products, LLC
• Original Assignee: Slmp, LLC (SLMP Holding LLC)
• Inventors: Imam, Syed Ashraf, Rees, Mark Lee
• Primary Examiner(s): Dauner, Joseph G.
• Application Number:
  US14/880,450
• Publication Number:
  US 20170059456A1
• Time in Patent Office: 1,352 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/4833 of solid biological material, e.g. tissue samples, cell cultures tissue in vivo A61B5/00; cell suspensions G01N33/48735
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

The disclosed embodiments include methods to form STC and STMC for use in determining presence of cancer, and methods to detect presence of cancer. In one embodiment, A portion of a STC is stained. The STC includes normal cells and cancer cells of a type of cancer co-cultured based on at least one cell culturing factors. The at least one co-culture factors includes a type of the cancer cells being cultured, a ratio of the cancer cells to the normal cells being co-cultured, seeding density of the normal cells and the cancer cells being co-cultured, a type of cell growth supplement used to facilitate culturing the cells, and a concentration of the cell growth supplement used to facilitate co-culturing the cells. The stained portion is observed to determine presence of one or more biomarker types that indicate presence of cancer.

Claims

1. A method to form a synthetic tissue control for use in determining presence of cancer, the method comprising:

preparing an approximately zero gravity environment for cell culture;

culturing cells comprising normal cells and cancer cells of a type of cancer in the approximately zero gravity environment and based on seeding density of the normal cells and the cancer cells being cultured; and

adding approximately 5 micrograms of DNase per milliliter of cell culture medium to facilitate un-clumping of the normal cells and the cancer cells during co-culturing of the cells.

2. The method of claim 1, wherein culturing the cells comprises co-culturing the normal cells and the cancer cells in a cell culture chamber configured to maintain a concentration of CO₂ and temperature inside the cell culture chamber.

3. The method of claim 2, wherein culturing the cells further comprises maintaining the cell culture chamber in a motorized rotating device operable to:

hold the synthetic tissue control; and

spin the cell culture chamber at a predetermined speed during co-culturing of the cells.

4. The method of claim 3, further comprising:

forming a homogenous core with un-clumped normal cells; and

invading the homogenous core with un-clumped cells of the cancer cells.

5. The method of claim 1, further comprising adding Fibronectin to improve contact between the normal cells and the cancer cells.

6. The method of claim 1, wherein culturing the cells comprises culturing the cells in the approximately zero gravity environment for a period of 8-12 days.

7. The method of claim 1, wherein culturing cells comprising normal cells and cancer cells comprises culturing normal cells and breast cancer cells.

8. The method of claim 7, wherein a ratio of the breast cancer cells to the normal cells is approximately 1:

99.8.

9. The method of claim 7, wherein the breast cancer cells are MCF.7 cancer cells.

10. The method of claim 9, wherein the seeding density of MCF.7 cancer cells is approximately 18,750 per milliliter.

11. The method of claim 1, wherein the seeding density of the normal cells is approximately 187,876 per milliliter.

12. The method of claim 1, further comprising staining the synthetic tissue control with a stain to observe one or more expressions of specific markers of the cancer cells.