Rat Mesenchymal Stem Cell Kit

Rat Mesenchymal Stem Cell Kit

Chemicon (Millipore)

Mesenchymal stem cells, also known as marrow stromal cells (1), are defined as a self-renewing population of adherent, bone-marrow-derived multipotent progenitor cells with the capacity to differentiate into several mesenchymal cell lineages. In defined in vitro assays, mesenchymal stem cells have been shown to readily differentiate into lineage-specific cells that form bone, cartilage, fat, tendon and muscle tissues (1, 2). Mesenchymal stem cells also provide support and maintenance for the other major stem cell population in the bone marrow, the hematopoietic stem cells (2).

Mesenchymal stem cells have historically been isolated based on the ability of these cells to form adherent cell layers in culture and the concomitant lack of adherence of other cells in the bone marrow stroma such as hematopoietic stem cells, adipocytes, and macrophage (1, 3). While this procedure results in enriched populations of mesenchymal stem cells, the resulting bone marrow derived cell populations are nonetheless, heterogeneous - comprised not only of mesenchymal stem cells, but also of committed lineage-restricted progenitors (1, 3). To aid researchers in the accurate identification and characterization of mesenchymal stem cells, CHEMICON® presents the Rat Mesenchymal Stem Cell Kit (Catalog No. SCR026).

CHEMICON®'s Rat Mesenchymal Stem Cell Kit provides ready-to-use primary mesenchymal stem cells isolated from the bone marrow of adult Fisher 344 rats along with a panel of positive and negative selection markers for the characterization of the mesenchymal stem cell population. Positive cell markers include antibodies directed against two cell-surface molecules (integrin b1 and CD54) (2, 4) that are present on mesenchymal stem cells. Negative cell markers include antibodies directed against two specific hematopoietic cell surface markers, (CD14, present on leukocytes and CD45, present on monocytes and macrophages) that are not expressed by mesenchymal stem cells (2, 3, 4). Mouse and rabbit immunoglobulins for the assessment of background staining are also included.

All of the antibodies provided in the kit have been tested and optimized for use in immunocytochemistry on rat mesenchymal stem cells. We recommend that CHEMICON®'s Rat Mesenchymal Stem Cell Kit (Catalog No. SCR026) be used in conjunction with differentiation assays (Catalog No. SCR020, Mesenchymal Stem Cell Adipogenesis Kit and Catalog No. SCR028, Mesenchymal Stem Cell Osteogenesis Kit) that demonstrate multipotentiality of the starting cell population.

For Research Use Only; not for use in diagnostic procedure.

Immunocytochemistry

Rat

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Stem Cell Kits

Mesenchymal Stem Cells

Cells/Isolation

• 1x10e6 viable Rat Mesenchymal Stem Cells: (Part No. 2004005) derived from adult female Fisher 344 rats, cryopreserved. Store in liquid nitrogen.
• Rabbit anti-Integrin beta1: (Part No. AB1952-20) One vial containing
• 20 mL polyclonal rabbit serum. Store at -20 °C.
• Mouse anti-CD54 (ICAM-1): (Part No. MAB2130) One vial containing 100 μL ascites monoclonal antibody. Store at -20 °C.
• Mouse anti-CD45: (Part No. 2003607) One vial containing 100 μL of
• 1 mg/mL monoclonal antibody. Store at -20 °C.
• Mouse anti CD14: (Part No. 2003608) One vial containing 10 μg of
• 0.1 mg/mL monoclonal antibody. Store at -20 °C.
• Mouse IgG: (Part No. PP54-100UG) One vial containing 100 μg of
• 1 mg/mL purified mouse IgG control antibody. Store at -20°C.
• Rabbit IgG: (Part No. PP64-100UG) One vial containing 100 μg of
• 1 mg/mL purified rabbit IgG antibody. Store at -20 °C

When stored at the recommended storage conditions (refer to Kit Components), components are stable up to the expiration date. Do not expose to elevated temperatures. Discard any remaining reagents after the expiration date. We recommend that the cells be used within ten passages.

1. Mesenchymal Stem Cell Expansion Media (DMEM-low glucose, without glutamine, 10% heat-inactivated fetal bovine serum (CHEMICON® Catalog No. ES-009-D), 2 mM L-Glutamine and (CHEMICON® Catalog No. TMS-002-C), 1X solution of penicillin and streptomycin (CHEMICON® Catalog No. TMS-AB2-C).

2. Accutase™ Cell Detachment Solution (CHEMICON® Catalog No. SCR005)

3. Chamber slides

4. Glass coverslips

5. Phosphate-Buffered Saline (1X PBS) (CHEMICON® Catalog No. BSS-1005-B)

6. Fixative (e.g. 4% Paraformaldehyde in 1X PBS)