This patent application is assigned to
The following quote was obtained by the news editors from the background information supplied by the inventors: "The invention relates to cell support compositions comprising a basement membrane extract isolated from cardiac or smooth muscle. The invention also relates to methods of using the cell support compositions for supporting cellular functions.
"Basement membranes are thin, continuous sheets that separate an epithelium from adjacent stroma and surround nerves, muscle fibers, smooth muscle cells and adipose cells. As shown in many species, basement membranes appear early during embryogenesis and cover the inner cell mass, suggesting a critical role for basement membrane in early development. All basement membranes contain members of 3 extracellular matrix families: laminins, collagen IV and proteoglycans, along with the glycoprotein, nidogen/entactin. The basement membrane is a nanoscale network of filaments that interacts to form a lattice. This network, in part, results from the assembly of independent networks of laminins and collagen IVs, joined together by the nidogens and other associated proteins.
"Basement membrane preparations have been used for cell biology research to provide a physiological environment promoting cell attachment, growth, and differentiation. Thus for culturing embryonic and other pluripotent stem cells in vitro, the basement membrane promotes cell growth and can be used to promote the differentiation of certain cell types.
"Among the most broadly employed basement membrane preparations is the basement membrane-rich matrix from Engelbreth Holm-Swarm (EHS) murine sarcoma, known by its trade name MATRIGEL.TM. (
"However, MATRIGEL.TM. has limited utility in many applications that arises from its origin as a mouse tumor or limits in the biological activity of the components that comprise it. Many products and processes for making these products require that the products or processes used to make the products be free of animal products. Clearly mouse-derived products are not suitable, for example, for the culture of human cells where the cells are grown to generate biologics products for human use such as antibodies or other proteins. The mouse-derived products are unsuitable for fabrication of tissue engineering scaffolds or other forms of implantation in humans due to concerns of passage of murine pathogens or elicitation of unwanted immune response. Beyond the concern of species difference is that of the composition itself. For example, MATRIGEL.TM. contains only laminin type I and alpha 1 and alpha 2 chains of collagen type IV. There are 6 chains of type IV collagen and 15 chains of laminin found in human cells, each assumed to impart specific biological functions to the basement membranes in which they occur.
"There are undoubtedly components of the basement membrane of each tissue that are unique to that given tissue, although this area of research still requires much greater elucidation. For example, the heart is known not to contain a specific population of stem cells that give rise to cardiomyocytes. Rather, the cardiac tissue contains specific signaling molecules like the chemokines, SDF-1, which recruits circulating stem cells to the heart upon damage. Likewise, the basement membrane composition of tissues like smooth muscle or cardiac muscle constitute a unique signature for each tissue."
In addition to the background information obtained for this patent application, NewsRx journalists also obtained the inventor's summary information for this patent application: "The invention relates to cell supports compositions comprising a basement membrane extract isolated from cardiac or smooth muscle tissue. The compositions comprise molecules from extracellular matrices including, but not limited laminin, collagen, perlecan, and nidogen/entactin. In some embodiments, the composition may further comprise additional heparan sulfate proteoglycans.
"The invention also relates to methods of using the novel compositions for supporting cellular functions, including but not limited to promotion of cell attachment, proliferation, differentiation, or maintenance of the phenotype of differentiated cells, or maintenance of the totipotency, pluripotency, multipotency of stem cells or progenitor cells through multiple culture passages in vitro
"The invention also provides methods of applying or injecting the cell support compositions of the present invention in a subject in need of treatment such treatment.
"The invention also relates to articles of manufacture comprising the cell support compositions of the present invention. For example, the invention provides for cell culture surfaces comprising the cell support compositions of the present invention.
BRIEF DESCRIPTION OF THE DRAWINGS
"FIG. 1 depicts the protein content of the cell support compositions of the present invention. Equivalent amounts of protein from 4 preparations of human heart basement membrane extract (HBME) from individual donors and MATRIGEL.TM. (MG) were resolved by electrophoresis on 5% SDS-polyacrylamide gels. The pattern of polypeptides in HBME was very reproducible and distinct from that of MATRIGEL.TM.. The majority of the protein in MATRIGELTM is present as laminin 1--the alpha 1 chain migrates at 400 kD and the beta 1 and gamma 1 chains co-migrate at about 200 kD. A number of the polypeptide bands in the region of 180-400 kD in the HBME samples reacted positively with monospecific antibodies against known laminin chains. Similarly, specific antibodies to type IV collagen (alpha 1 and alpha 2 chains), type I collagen, nidogen/entactin and perlecan were able to bind to the HBME. Nidogen was also present. Tandem mass spec sequencing analysis provided further confirmation of the identity of these polypeptides and identified the presence of several non-basement membrane cytoskeletal proteins, including titin protein, myosin 7 and actinin.
"FIG. 2 depicts the attachment of cells to the cell support composition of the present invention. 20,000 of each cell type was allowed to attach to a surface coating of 10 .mu.G/cm.sup.2 for 1 hour at 37.degree. C. in serum free media. The Table in this figure shows the ratio of attachment to the cell support compositions of the present invention relative to MATRIGEL.TM.. For stem cells and fibroblasts, the cell support compositions of the present invention promote roughly twice the level of attachment as compared to MATRIGEL.TM.
"FIG. 3 depicts the growth and maintenance of plasticity of pluripotent stem cells cultured on the cell support compositions of the present invention and on MATRIGEL.TM.. Either the cell support compositions of the present invention or MATRIGEL.TM. was coated at 17 .mu.G/cm.sup.2 were tested for the ability to support the growth and prevent the unwanted differentiation of embryonic and induced pluripotent stem cells in serum-free, chemically defined medium. Mouse embryonic stem cells (D3), mouse induced pluripotent stem cells (W5) or human induced pluripotent stem cells were plated and passaged 2 times on the indicated substrates. Proliferation rates and colony morphology was comparable. When the human induced pluripotent stem cells were stained at second passage using a live cell marker for pluripotency (Tra-1-60, stain alive, Stemgent) the green FITC signal indicates the cells were still in a pluripotent state."
URL and more information on this patent application, see: Ogle, Roy. Basement Membrane Compositions and Applications Thereof. Filed
Keywords for this news article include: Antibodies, Laminin, Peptides, Treatment, Immunology, Amino Acids, Blood Proteins,
Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC
Most Popular Stories
- Photo ID Required for Unemployment Benefits
- Software Writers Sought in Indiana
- Ukraine Crisis Limits Losses in Gold, Silver
- Can GOP Dodge Immigration Bullet?
- How Past Mistakes Will Drive Ukraine's Future
- Tech Firms to Increase Hiring for 4th Year in a Row
- Job Fair for S.C. Grads
- Chiquita, Fyffes to Form Top Banana
- Millennials Favor Saving Over Investing: UBS
- Big Earthquake Rumbles Northern California