Technology: Cell system development to isolate and expand useful cell types from human and animal tissues for research and therapeutic applications. This involves development of cell separation technology to identify and isolate specific cell types from various tissues and development of growth media for such specific cell types. These cells are used as primary cells for various applications or immortalized to obtain a continuous source of cells for drug discovery, high throughput screening (HTS), drug metabolism, toxicology, and cell therapy applications. The immortalized cell lines are isolated to ensure retention of almost all of the biochemical properties of corresponding cognate primary cell types.

Clonexpress is currently working on the following areas of development of cell systems for research and therapeutical applications.

Human Brain Derived Cell Types:

Expansion of Human Neuronal Precursor Cells (HNPC) and Development of Human Neuronal Cell Lines

Clonexpress, Inc., has developed proprietary processes to isolate and expand HNPCs from human brain for several passages. Using this method, it has been possible to expand HNPCs more than thousand fold. The HNPCs can be maintained for 6 passages or more in the proprietary media containing fetal bovine serum (FBS) and growth factors. The cells can be made to differentiate into mature neurons by plating them in both serum free medium and medium containing serum on poly L-lysine (PLL) coated dishes  with growth factors and dibutyryl cAMP or NGF. Extra cellular matrix components, such as laminin, further accentuates differentiation into neurons. The neurons express several neuron specific markers (MAP 2, beta tubulin III), and form extensive processes. They also have electrical activity. HNPCs may also respond to other differentiation promoting agents, such as retinoic acid and other neurotrophic factors, in giving rise to specific types of neurons. Cells in serum containing differentiation medium can be maintained for several weeks.  HNPCs are immediately available for research. (Cat no: HNPC 010)

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Human Neuronal Precursor Cell Line (HNPCL):

Clonexpress is also developing HNPC lines from human brain tissue. Some of the neuronal cell lines are truly precursor cell lines, and they express both GFAP and neuron specific markers (MAP2ab). In this regard, they are not fully committed cells, and they have the potential to develop either into astrocytes or neurons by altering culture conditions. The precursor cell lines develop into mature neurons when plated with dibutyryl cAMP or NGF both under serum free and serum containing growth conditions on PLL coated dishes with and without laminin, whereas the same cells develop into astrocytes when plated with serum under similar conditions but leaving out growth factors.

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Microglial cells and cell lines (MG):

Clonexpress has developed a proprietary process to amplify human brain derived microglial cells for 6-10 population doublings using a specific medium developed for these cells. The microglial cells express a HLA class II antigens following treatment with interferon gamma, CD68, CD14, esterase, and other markers. The microglial cells do not express GFAP (astrocyte marker) or neuron specific markers. Human microglial cells are immediately available for research applications. Clonexpress is also developing microglial cell lines from human brain tissue. The microglial cell lines express all the functions similar to the primary microglial cells. They secrete cytokines in response to pro inflammatory cytokines. Some of these cell lines can be grown in continuous culture for an extended period of time compared to primary microglial cells.  These cell lines can be used as research tool to study HIV replication and drug development to block neurodegenerative disorders in AIDS and other diseases. ( Microglial cell catalog No: HMG 030)

Astrocytes and astrocyte cell lines:

Primary human astrocytes are immediately available for research. These cells are supplied with a proprietary growth factor supplement and are guaranteed to grow for a minimum of 8-10 population doublings. These cells express GFAP and do not differentiate into neuron like cells when plated on PLL coated plates in serum containing medium with growth factors and dibutyryl cAMP or NGF. Clonexpress is also developing human astrocyte cell lines for HTS drug discovery applications. (Astrocyte cell catalog No: HAST O40)

Matched pair of human neuronal precursor, astrocyte, and microglial cell lines:

Clonexpress is developing human neuronal precursor, astrocyte, and microglial cell lines from the same donor. Such matched pairs of cell lines will enable coculture of them in order to understand mechanism of neuronal differentiation, and neuronal cell death under the influence of various agents. These matched cell systems will be useful to develop drugs for neurodegenerative disorders as well as ischemia induced neuronal cell death.  Matched cell lines of all of these cell types are developed for drug development applications.

Brain cells under development:

Human Dopaminergic Neurons (DAN):

Clonexpress has successfully developed proprietary methods to similarly expand dopaminergic neuronal precursors. Preliminary results indicate that such cells can specifically be isolated and expanded to a moderate extent. Currently, Clonexpress is improving culture conditions to obtain a significant expansion of dopaminergic neurons in vitro. This has important therapeutical potential for Parkinson’s and other disorders, where dopaminergic neuronal precursor cells expanded in vitro can be used to treat several patients. Clonexpress is also developing dopaminergic neuronal cell lines. A few cell lines that have been selected are able to grow in tyrosine free medium, and express tyrosine hydroxylase. Primary DAN cells are immediately available for research (Catalog No: DAN 020)

Neuronal precursor cells and cell lines from Down’s syndrome:

Clonexpress is developing HNPCs from Down’s syndrome tissue samples. Primary Down’s HNPCs are available for research. Down’s HNPC cell lines and Down's syndrome DAN cell lines are expected to be available for evaluation and licensing in the future.  Clonexpress is also developing microglial cells from Down's syndrome. These cells are expected to be available shorthly. Neuronal precursor cells and other brain derived cells from Down's syndrome may be useful to investigate some aspects of Alzheimer's disease.

How to obtain HNPCs and other brain derived cells:

Contact Clonexpress about obtaining HNPCs and other CNS cells along with the medium to maintain these cells for research. These cells are only for research applications, and not intended for any therapeutic use in humans. User of cells from Clonexpress also agrees that he or she will not sell or distribute cells, subcultures, and fractions thereof to any third party without obtaining a prior written permission. The cells are sold to the researcher for only their own research.

Primary cells are normally supplied as live cell suspension or grown in a T-25 tissue culture flask along with cell specific growth factor supplement sufficient to make 500 ml of culture medium. Clonexpress will also supply a detailed instruction of handling procedures along with the cells. Cells can also be obtained in cryopreserved format, and such cells are supplied in dry ice. Dry ice shipments require additional fee in addition to the normal shipping cost, which is added to the invoice.

HNPCL, astrocyte, and microglial cell lines are expected to be available for evaluation and licensing. These cell lines are given for evaluation after executing a Technology Evaluation Agreement (TEA) and a confidentiality agreement. The cell lines are then licensed involving both site specific and time specific terms. Contact Clonexpress for further information about licensing these cell lines:

Clonexpress, Inc.,

504 E.Diamond Ave, Suite G

Gaithersburg, MD 20877

Ph: 301-869-0840; Fax: 301-963-4214

Clonexpress strives to be a provider of all human CNS cells to research community so that they can focus on their discovery efforts. Clonexpress is also developing human CNS cells for therapeutic applications.

Other Cell Types Under Development:

Human hepatocyte,  and pancreatic beta cell lines:

Clonexpress has developed a proprietary technology to immortalize human hepatic precursor cells using liver specific selection scheme that allows the growth of only hepatic cells.  Clonexpress is also working on the development of human pancreatic beta cell lines that secrete insulin in a highly glucose regulated fashion. Clonexpress is currently characterizing these cell lines.

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Other Services:

Custom development of Cell lines:

Clonexpress can custom develop cell lines using primary cells provided by the client or isolated from tissues procured by Clonexpress. This will include optimization of growth conditions for the cell type and development of several clones of immortalized cell lines. The cell lines will be characterized as per the specifications of the client. Any special reagents necessary for the characterization of cell lines shall be provided by the client. Please send inquiries to Clonexpress.

Gene Transfer:

Clonexpress can also undertake transfer of genes into cell lines on custom basis. Both the genes and the cell lines into which such genes need to be stably introduced shall be provided by the client. Clonexpress will isolate several clones and characterize the clones for expression of exogeneously introduced genes using procedures provided by the client.

Warranty

Primary cells supplied by Clonexpress are negative for HIV, hepatitis B and C. The cells are also negative for bacteria, fungi, and mycoplasma. All characteristics of the cells and cell lines provided by Clonexpress are not fully understood and its use may involve risks or dangers that are not known or fully appreciated. Therefore, cells supplied by Clonexpress should be handled only by personnel skilled in proper cell culture techniques and laboratory procedures. The cells are being provided without any warranty of any sort other than the guarantee for the expression of functions described herein. The customer will use the cells provided by Clonexpress in compliance with all laws and governmental regulations and guidelines applicable to the use of tissue culture cells and human tissue.

For further information and to order cells contact info@clonexpress.com