Stem Cell Protocols

Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes, and osteocytes (1, 2). MSCs are phenotypically characterized as CD34^-, CD105⁺, and CD90⁺ cells. This protocol describes the expansion of human MSCs using the StemXVivo™ MSC Expansion Media (R&D Systems, Catalog # CCM004).

Please read the protocol in its entirety before starting.

This protocol provides a basic guide for the preparation and fixation of stem cells on glass coverslips for downstream applications such as immunocytochemistry. As many cultured cell types do not adhere well to glass coverslips, adhesion of cultured stem cells to glass can be enhanced by adding a thin layer of substrate on which the stem cells are normally cultured to the coverslip. Refer to the protocols outlined here for more detailed instructions.

Ex vivo expanded neural stem cells serve as excellent tools for researchers studying neural development and neurological disorders. Ready-to-use primary cortical stem cells, isolated from E14.5 Sprague-Dawley rats (Catalog # NSC001), can be grown in monolayer or as neurospheres as described here. These cells retain capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes.
Please read the protocol in its entirety before starting.

Mesenchymal stem cells (MSCs) are functionally defined by their capacity to self renew and their ability to differentiate into multiple cell types including adipocytes, chondrocytes, and osteocytes. This protocol describes a technique to promote the osteogenic differentiation of human and mouse MSCs.

Please read the protocol in its entirety before starting.

The colony forming cell (CFC) assay, also referred to as the methylcellulose assay, is an in vitro assay used in the study of hematopoietic stem cells. The assay is based on the ability of hematopoietic progenitors to proliferate and differentiate into colonies in a semi-solid media in response to cytokine stimulation. The colonies formed can be enumerated and characterized according to their unique morphology.

Please read the protocol in its entirety before starting.

This protocol must be read in its entirety before using Catalog # NSC002 and AR003.

The colony forming cell (CFC) assay, also referred to as the methylcellulose assay, is an in vitro assay used in the study of hematopoietic stem cells. The assay is based on the ability of hematopoietic progenitors to proliferate and differentiate into colonies in a semi-solid media in response to cytokine stimulation. The colonies formed can be enumerated and characterized according to their unique morphology.

Please read the protocol in its entirety before starting.

If culturing in the absence of a feeder cell layer is desired, human embryonic stem (hES) cells can be maintained using Mouse Embryonic Fibroblast Conditioned Media (Catalog # AR005). The protocol below has been used with the BG01V line of hES cells. Please note that other hES cell lines may require modifications of this protocol. Optimal culture conditions must be determined by the investigator for each hES line.

Please read the protocol in its entirety before starting.

Human embryonic stem (hES) cells can be maintained on a layer of mitotically inactivated feeder cells such as irradiated mouse embryonic fibroblasts (iMEF, Catalog # PSC001). The protocol below has been used with the BG01V line of hES cells. Please note that other hES cell lines may require modifications of this protocol. Optimal culture conditions must be determined by the investigator for each hES line. Please read the protocol in its entirety before starting.