Cell Biology

 ESTABLISH PRIMARY CELL CULTURES         IMMORTALIZE  & GROW  B-CELLS           EXPAND TUMOR CELL LINES       CELL BASED ASSAYS 

KTI offers a wide-range of cell culture services associated with the establishment, expansion, cryo-preservation and storage of normal and atypical cell lines. We provide a highly effective and efficient arrangement for your experimental needs of cultured cells. Cultures are established to meet your specifications for animal source, dishes for plating, cell density and any other specified parameters. Cultures are established with a few days notice for the first time and on specified delivery days thereafter. KTI technical staff works closely with the investigators to identify and comply with their specific needs. Following are the examples of such cell culture operations at KamTek Inc.

 1. PRIMARY CELL CULTURES 
 ---PRIMARY CULTURES OF HUMAN PBMCs

  • Isolate Peripheral Blood Mononuclear Cells (PBMCs)
  • Cultures of Activated B-cells, T-cells & Macrophages
  • Infect Cultures with a Virus e.g. HIV,
  • EBV transformation of B-cells
  • Establish fibroblast and epithelial cell cultures from excised tissues
  • Treat Cultures with other Agents to Study Cell Response
  • Cryo-Preserve and Store PBMCs in Liquid Nitrogen
  • Ship Frozen or Viable Cultures Worldwide

---ANIMAL PRIMARY CULTURES

  • Activated B-, T-cells or macrophages from PBMCs, spleen or thymus tissue
  • Generation of B- and T-cell hybridomas
  • Neurons, Astrocytes,
  • Epithelial cells,
  • Adipose cells, and Fibroblasts
  • Cryo-preservation for long term cryo-storage of cells

 

 

 

2. ESTABLISH B-CELL LINES BY EBV TRANSFORMATION

Buffy coat isolated from human blood sample received from a donor is transformed with EBV, and in 4-8 weeks, continuously growing culture can be established as a resource of:·

  • Genetic Material Of B-Cells·
  • Amplified DNA· and
  • Human Monoclonal Antibodies

KTI technical staff works closely with the investigators to identify and comply with their specific needs. For example, depending on the needs of the project, EBV transformation may be set up with:

  1. Relatively fewer PBMCs keeping remaining as a back-up cryo-preserved stock,
  2. Cryo-preserved PBMCs from a client, 
  3. Cord blood cells, or 
  4. All available fresh PBMCs for bulk transformation to screen for human Monoclonal antibody productions followed by cloning steps.

Transformed cells are grown and cryo-preserved for future use. In addition, cell line growth can be continued to harvest cells for DNA isolation. Cell stocks are always tested for lack of mycoplasma contamination and can be validated for specific characterization before cryo-preservation. Alternatively, bulk cultures prepared can be shipped to client for testing. Cryo-storage services are provided at clients request. 

3. TUMOR CELL LINE EXPANSIONS
to:
  • Harvest Cells or Secreted Products
    • recombinant gene products
    • growth factors
    • necrosis factors or
    • microbial agents such as HIV
  • Re-Clone and Grow B-Cell Hybridomas
  • Study Tumor characteristics
  • Cell line procurement from ATCC to grow for experiments
  • Long Term Storage for later reference

All cultures expanded at KTI go through MYCOPLASMA TESTING at least at two stages-- at the mid point of expansion and when culture is fully expanded.

4. CELL BASED ASSAYS 

  • Microbial Infections by In-situ Hybridization assays
  • Immunofluorescence assays (IFAs) on Tissues & Cells
  • Immune-function Assays
  • Cytotoxicity Assays
  • Cell line identification
  • Support Transfection Studies
  • Induce Differentiation: monocyte to macrophages; fibroblasts into adipose cells
  • Plate Cultures
  • Cultures on Multi-spot or Multi-chamber Slides