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Extracellular Matrix (ECM) Coated Plastic Ware

Plastic Vessels

Coated with Extracellular Matrix (ECM) Simulate in‑vivo Conditions

 

ECM ‑ The Natural Substrate

One of the drawbacks in growing cells in vitro using conventional tissue culture techniques is that the cells rest on plastic rather than on their natural biological support.  This natural support is a complex network of macro­molecules known as the extra­cellular matrix or ECM.  ECM holds cells and tissues together and provides a highly organized lattice within which cells can migrate and interact with each other.  The matrix plays an active and complex role in regulating the behavior of cells that are in contact with it, influencing their shape, migration, proliferation and metabolic functions. In contrast, cells grown on plastic lose many of their natural differentiated properties due to the lack of interaction with ECM.

 

ECM Technology

Growing cells outside the body requires creating conditions that mimic as closely as possible the situ­ation inside the body. Thus, it is essential that anchorage‑dependent cells be grown in contact with a natural extracellular framework.  Towards this end a new technique for coating tissue culture dishes with a naturally produced basement mem­brane‑like extracellular matrix (ECM) has been developed.

 

During the search for an appropriate tissue culture substratum, it was found that cultured endothelial cells, either vascular or corneal, secrete a large amount of extracellular matrix on to the plastic surface.  This material is similar in organization and chemical composition to naturally occurring basement membranes upon which cells migrate, proliferate, and differentiate in vivo.  When the endothelial cells are removed they leave the underlying ECM intact and firmly attached to the plastic.

 

Cells placed in contact with ECM attach rapidly, exhibit high plating and cloning efficiencies, proliferate rapidly, reach a high saturation density, exhibit lower requirements for serum and added growth factors, respond better to physiologically occurring hormones, express differentiated functions, have longer life span, undergo flattening and morphological changes, and have better plating consistency.

 

In order to improve cell attachment and growth, isolated matrix components such as collagen, laminin and fibronectin have been used to coat tissue culture plastic.  However, it has been found that cells placed in contact with ECM adopt growth characteristics, morphological appearance and biological responses which are not expressed when the same cells are maintained on artificial substrata (plastic, glass), even if coated with isolated constituents of ECM such as purified collagen or glycoproteins.  The use of naturally produced ECM, assures that the various matrix components will be found in their natural configuration and proportion.

 

Among the cells types showing a favorable response to ECM are human, bovine and other origin.

 

Applications of Extracellular Matrix to Culturing of Epithelial Cells

Epithelial cells are the principal producers of most hormones, enzymes and other secretory proteins.  Epithelial cells also constitute the most common target cells of human cancer.  Growth of epithelial cells is therefore of prime importance for various research, clinical and industrial purposes.  The main difficulty in obtaining pure and actively growing epithelial cell cultures is that the epithelial cells tend to be over­grown by stromal fibroblasts.  Hence, the main requirements for growing primary and early passage epithelial cells are growth conditions that support epithelial out-growth and suppress proliferation of adjoining fibroblasts.

 

ECM‑coated plastic vessels with serum‑free medium enable a higher rate of success in growing normal and malignant human epithelial cells from biopsy specimens.  The higher rate of success is due to  better plating efficiency and active epithelial cell proliferation in a serum‑free medium which suppresses the growth of fibroblasts.

 

ECM induces changes in cell shape not observed in cells grown on plastic or isolated components of the ECM.  Cells which for different reasons do not flatten or spread on plastic do so rapidly on ECM.  These cells are more receptive to hormones and naturally occurring factors.

 

ECM/Serum Free for Hormones, Enzymes, Growth Factors and Other Cellular Products

Hormone Secretion Research

ECM‑coated plastic vessels with serum‑free media support the maintenance and normal function of hormone secreting cells such as pancreatic islet cells, hepatocytes  pituitary cells, granulosa cells, etc.  The ability to culture cells from endocrine organs makes it possible to study control mechanisms of hormone production.  This is essential for both basic studies on the control of mechanisms of hormone production and secretion and for industrial applications requiring inducement of maximal hormone secretion. Demands for cellular products such as hormones, growth factors and enzymes are increasing for both basic and industrial applications.

 

Secretion of Cellular Products

The ECM/serum‑free medium combination promotes research possibilities on various cellular products due to the following multiple effects:

 

Hormone Response Research

ECM effects cell shape and hormone responsiveness.  It is thought that the basal lamina or ECM exerts its permissive effects on cell proliferation and differentiation by modifying cell shape to make it more responsive to hormones and other naturally occurring factors.  As expected the cells do not respond when maintained on artificial substrata or isolated components of the ECM.  Growing hormone‑responsive cells in serum‑free environment will eliminate the effects of other hormones and growth factors which are present in serum, and will allow the study of the isolated effects of a specific growth factor or hormone.

 

Hormone Secreting Cells

Pituitary cells: the pituitary gland secretes a variety of hormones controlling several important functions, ranging from growth and differentiation to reproduction.  Of major interest is the production of growth hormone and thyroid stimulating hormone (TSH). Synthesis and secretion of TSH are regulated by the levels of thyroid hormones in the circulation and by the secretion of thyrotropin releasing hormone (TRH) by the hypothalamus.  Plating adult rat pituitary cells on ECM enabled the development of pituitary epithelial cell monolayers that retain, for about two weeks in culture, their in vivo hormone secreting and responding characteristics.  With the aid of ECM‑coated plastic vessels it is possible to further clarify the molecular and control mechanisms of TSH and growth hormone production by the pituitary gland.

 

Pancreatic beta‑islet cells: when plated on ECM, beta‑islet rat cells respond to glucose and secrete insulin to a higher extent than when coated on plastic.

 

Granulosa cells: ECM enables the growth of rat, bovine and human granulosa cells, which otherwise do not grow well and lose their specific properties.  ECM has been shown to promote cell communication and induce higher secretion of steroid hormones by granulosa cells.

 

Hepatocytes: ECM has been shown to support the attachment and monolayer formation by normal hepatocytes.This will enable studies on the production of specific liver enzymes, hormones (transferin) and proteins.

 

Biotechnical Applications

Yield and differentiation:  the maintenance and growth of differentiated cells on ECM is expected to promote a high yield of various hormones and growth factors in tissue culture.

 

Purification: growth of cells in serum‑free media will facilitate the purification of various cellular products that are secreted into the medium.  Purification will be relatively simple due to the absence of serum proteins.

 

Production: large‑scale growth of cells on ECM can be performed in bulk cell culture vessels coated with ECM, or on ECM‑coated microcarriers. Using these techniques, continuous rather than batch processes can be developed.

 

Growth factor secretion: growth factors may be produced in better yields by human cells cultured on ECM rather than on plastic and can then be purified and used for research and clinical applications.

 

Other Applications

In vitro toxicological testing and drug screening ‑ alternative to laboratory animals: the ability to grow primary cells in vitro for drug screening and toxicological testing may offer an alternative to the use of laboratory animals.  The growth of cells on ECM in serum free medium may reduce the cost and simplify the procedure of studying the effect on cells of single drugs, drug combinations and hormones or where a single component is being tested at a time.

 

Neurobiology: ECM has been shown to support the attachment and maintenance of neurons from  various sources and to promote the outgrowth and directed elongation of neurites.  It has also been shown to induce the proliferation and network formation of adult rat brain oligodendrocytes maintained in a serum‑free medium.

 

Novamed Ltd  produces plastic tissue culture ware coated with ECM from bovine corneal cells. We grow the cells in several different types of vessels (plates, flasks, microtiter plates, etc.), and after they reach confluence, the cells are removed leaving the ECM behind.

 

Following types of Extracellular Matrix (ECM) Coated  Plastic Ware are available for ordering:

 

ITEM

Packaging (unit/Pack)

Catalogue

 No.

TISSUE CULTURE DISHES 35 Mm

5

E-TCP-35

TISSUE CULTURE DISHES 60 mm

5

E-TCP-60

TISSUE CULTURE DISHES 90 mm

5

E-TCP-90

TISSUE CULTURE FLASKS 25 cm2

5

E-TCF-25

TISSUE CULTURE FLASKS 80 cm2

5

E-TCF-80

Microtiter 96-Well Plate

1

E-TCMT-F

4-Well Culture Plate

1

E-TCMW-4

6-Well Culture Plate

1

E-TCMW-6

12-Well Culture Plate

1

E-TCMW-12

24-Well Culture Plate

1

E-TCMW-24

COVERSLIPS (ROUND, 22 mm)

5

E-TCCS-P22

FOUR 13mm Coverslips In 4-Well Plate

1

E-TC-IF-13

EIGHT 12mm FILTERS IN 24-WELL PLATE

1

E-TC-M-12

Eight Well Lab-Tek Chamber Slide

1

E-LT-8

FOUR 30mm FILTERS IN 6-WELL PLATE

1

E-TC-M-30

 

ECM Plastic Ware are Used like Regular Culture Vessels

ECM‑coated plastic vessels are shipped by express air mail and should  be stored at 4oC upon arrival.  Expiration date is one year from completion of production.  ECM‑coated dishes, flasks and slides are handled and used exactly like regular tissue culture plastic ware.

 


     FOR FURTHER INFORMATION EMAIL US OR CALL: +972-2-6781861


 

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