Research Tools : Cell lines


Multicolor Reporter Cells for Detecting and Quantifying HIV-1

UW–Madison researchers have developed highly sensitive and specific reporter cell lines suitable for automated detection of HIV in a microfluidic platform. The multicolor fluorescence-based cell readouts respond robustly to HIV-1 infections, and are useful for tracking the spread of HIV-1 infection and ideal for implementation in an automated Q-VOA assay.

The reporter cells are based on coupling fluorescent markers turned ‘on’ in response to HIV infection to markers turned ‘off’ by the virus. Calculating net ‘on/off’ ratios over time, relative to standards, allows for high sensitivity and favorable signal-to-noise. The ability to amplify response signals with minimal background and without the need for chemical substrates represents a significant improvement over existing green fluorescent protein (GFP) or chemiluminescence-based single reporter lines.

Adapted Rhinovirus C for Maximum Virus Yield

Building on their work, the researchers have now developed a mutated RV-C strain that induces strong cytopathic effect and replicates vigorously in the HeLa-E8 cells, yielding more than a log higher level of infectious rhinovirus particles compared to the parental clinical isolate.

New Viral Propagation Method Yields Insight Into Childhood Asthma

UW–Madison researchers have developed an efficient, cost-effective method of propagating RV-C. They discovered that human cadherin-related family member 3 (CDHR3) is the receptor for RV-C and allows cell lines normally unsusceptible to HRV-C infection to support virus binding and replication.

To create cell lines capable of efficiently growing RV-C, the researchers modify the host cell so it expresses an effective amount of the CDHR3 receptor. This method enables high-throughput, large-scale production of RV-C, which in turn enables critical basic and applied research regarding this understudied pathogen.

Multipotent Lymphohematopoietic Progenitor Cells

UW-Madison researchers have developed a population of cells derived from hES cells that comprise unique, multipotent lymphohematopoietic progenitors. The cells, which were obtained from co-culture of hES cells with OP9 stromal cells, are CD34 and CD43 positive, but CD45 and lin negative. These cells express gene profiles characteristic of definitive, multipotent hematopoietic progenitors and are capable of differentiating into lymphocytes, myeloid cells, erythroid cells and megakaryocytes. They are no longer capable of differentiating into non-hematopoietic lineages. If the cells are cultured until they express CD45, a cell surface marker found in hematopoietic progenitor cells derived from somatic (adult) stem cells, they lose the ability to produce differentiated cells of B lymphoid lineages.

Stable Cell Lines Expressing hERG1a and 1b

UW-Madison researchers have developed a line of cultured mammalian cells that express both the HERG1a and 1b subunits. In addition, they developed the only known antibody specific for the HERG1b isoform (which cross-reacts with ERG1b of rat and canine as well), and a line of cultured mammalian cells that express HERG1b but not HERG1a.

Common Variants of the Sodium Channel Alpha Subunits

UW-Madison researchers have identified four groups of SCN5A variants that represent the most common SCN5A variants in humans. The researchers found that none of the three previously known SCN5A clones represented a common sequence for SCN5A. The new SCN5A variants, on the other hand, have been observed in hundreds of individuals. Thus, they provide the true reference or background sequences for evaluating the normal functions of the sodium channel. They are also the true reference sequences against which the effects of various SCN5A mutations should be judged.

The researchers have put the four full constructs into appropriate expression vectors. They also have created mammalian cell lines that stably express each of the channels.

Expressible cDNA Encoding Human Calcium-Activated Potassium Channels

UW-Madison researchers have isolated cDNAs encoding the human and mouse homologs of the Drosophila calcium-activated potassium channel (BK) gene, called slowpoke. These cDNAs can be incorporated into different vectors for expression in cell lines or in Xenopus oocytes.

Cell Line Stably Expressing KvLQT1 and minK

UW-Madison researchers have developed an HEK 293 cell line that stably expresses KvLQT1 and minK. Since unintended block of potassium channel activity by drugs can cause an acquired form of long QT syndrome, which leads to potentially fatal arrhythmias, this system provides an important screening tool for drugs in development.

Cell Line Stably Expressing the Human Heart Sodium Channel Beta1 Subunit

UW-Madison researchers have developed a stably transfected cell line expressing the beta1 subunit of the human heart sodium channel. The beta1 subunit was first cloned in the early 1990s. The researchers have now re-cloned this subunit and created a cell line that permanently expresses it.

Human Heart Sodium Channel Beta1 Subunit (SCNB1)

The beta1 subunit of the human heart sodium channel is now available as a biomaterial through WARF. SCNB1 was first cloned in the early 1990s. UW-Madison researchers have now re-cloned this subunit and created a cell line that stably expresses it.

Immortalized Cells from the Mouse Urogenital Sinus and Adult Mouse Prostate

A UW-Madison researcher has developed biological material that includes immortalized murine mesenchymal cells from the urogenital sinus and stromal cells from the lobes of the adult prostate. The cells were isolated from the INK4A transgenic mouse, which lacks genes that normally enforce senescence. A construct that expresses lacZ was included in these cells to allow their identification when they are grafted into mouse tissues.

HERG-1 Transfected HEK 293 Cell Line B

UW-Madison researchers have cloned the full-length cDNA of the HERG-1 cardiac potassium channel gene into human embryonic kidney (HEK 293) cells, allowing expression of these channels in an experimental system. Since unintended block of HERG channel activity by drugs can lead to potentially fatal arrhythmias, this system provides an important screening tool for drugs in development.

Cell Line for Evaluating Influenza Virus Sensitivity to NA Inhibitors

A UW-Madison researcher has developed a cell line that is capable of universally monitoring the sensitivity of human influenza virus isolates to NA inhibitors. This Madin-Darby canine kidney (MDCK) cell line has been modified so it overexpresses the human β-galactosidase α2,6-sialyltransferase I (ST6Gal I) gene.

Several influenza virus isolates were tested in this cell line. The sensitivity of the viruses to an NA inhibitor correlated with the sensitivity of viral sialidase to the compound, demonstrating the potential utility of this cell line for detecting viruses that are resistant to NA inhibitors.

Methods and Materials for Assaying Non-SCD1 Isoforms

UW-Madison researchers have developed useful tools for the discovery of SCD inhibitors. The invention consists of cDNAs for murine SCD2 and SCD3 and human SCD5, along with a stable mammalian cell line and yeast strain that express human SCD5. In addition, the invention includes SCD2, and SCD3 knockout and transgenic mice, as well as the targeting constructs used to generate the SCD1 and SCD3 transgenic mice.

HERG CDNA with Native PolyA Tail

UW-Madison researchers have developed the full-length cDNA of HERG1a. This cDNA has been cloned and expressed in Xenopus oocytes, human embryonic kidney cells (HEK293) and Chinese hamster ovary cells. It is similar to a previously cloned cDNA, except that a polyadenylation signal has been removed from its vector sequence. The removal of this sequence does not affect HERG expression levels in heterologous systems.

Cell Line Stably Expressing Human Cardiac Ion Channel Common Polymorphism, K897T

UW-Madison researchers have developed a new human embryonic kidney (HEK 293) cell line that stably expresses the K897T polymorphism of the HERG potassium channel. The K897T polymorphism is the most common HERG polymorphism and is present in 25 to 30 percent of humans. Experiments suggest that carriers of this polymorphism may have a subtle but discernable phenotype that is distinct from the wild-type phenotype.

Clonal Lines of Human Cervical Epithelial Cells That Harbor Human Papillomavirus Type 16

UW-Madison researchers have developed clonal lines of human cervical epithelial cells that stably harbor the double-stranded, circular human papillomavirus type 16 (HPV16) genome. One set of cell lines contains HPV16 DNA as an extrachromosomal nuclear plasmid. These cell lines are useful for studying aspects of the viral life cycle, including host and viral requirements for viral DNA replication and transcription. The other cell lines comprise HPV16 DNA integrated into their genome, similar to what is observed in cervical cancer. Both sets of cell lines were subcloned from the same parent population, and therefore are closely matched.

New Cell Lines Containing Stably Expressed KATP Pores for High Throughput Screening

UW-Madison researchers have created two cell lines in which a KIR6.1 or KIR6.2 gene is stably expressed in COS1 cells.  Because the KATP pores are stably expressed, they should provide an improved tool for studying SUR isoforms and splice variants.  COS1 cells also have no background SUR.

These cell lines potentially could be used to identify novel compounds for cardiovascular disease, including hypertension, heart failure and myocardial protection from ischemia.  They also could be useful in the development of therapies for other conditions, such as type II diabetes.

Mouse Strains and Cells Useful to Investigate Tissue-Specific Roles of Stearoyl-CoA Desaturase Isoforms

A UW–Madison researcher has developed several mouse strains and cells useful in the study of SCDs.  To understand the tissue-specific role of SCD1 and other SCD isoforms in disorders such as obesity, diabetes, cancer, inflammation, atherosclerosis and metabolism, the researcher generated mice with tissue-specific knockouts of SCDs.  He also developed transgenic mice in which SCDs can be overexpressed in specific tissues.

Reprogrammed Stem Cell Line for Research: IISH3i-CB6

UW–Madison researchers have developed a reprogrammed iPSC line called IISH3i-CB6. Their method generates iPSCs free of transgene and vector sequences from human bone marrow and cord blood mononuclear cells using non-integrating episomal vectors.

Reprogrammed Stem Cell Line for Research: IISH4i-CBT4

UW–Madison researchers have developed a reprogrammed iPSC line called IISH4i-CBT4. Their method generates iPSCs free of transgene and vector sequences from human bone marrow and cord blood mononuclear cells using non-integrating episomal vectors.

Thiazovivin was added to improve reprogramming efficiency when creating this line.

Cryopreserved Head and Neck Cancer Tumorgrafts and Related Cell Strains

UW–Madison researchers have created a series of tumorgrafts from head and neck cancers, as well as cell lines from a subset of these tumorgrafts. Tumorgrafts are established by grafting human tumor samples directly onto mice and then passaging them onto other mice without ever placing them in tissue culture. They are expected to be more representative of actual human tumors than tumor xenograft mouse models.

Bovine Macrophage Cell Line

UW–Madison researchers have developed the first proliferating bovine cell line proven to have features of monocytic/macrophage lineage. The new cell line is useful for bovine disease research, such as studying host-parasite relationships from an immunological perspective.

The new line was discovered when some of the primary macrophage cells isolated from bovine blood samples began spontaneously proliferating under standard cell culture conditions. They were identified as a well-differentiated macrophage cell line based on morphological features and the presence of a macrophage surface marker, which was found in 99 percent of the cells. They also tested positive for markers confirming their monocytic and mesenchymal origins, respectively.

Cell Lines for High Throughput Screening of Estrogenic Compounds

A UW–Madison researcher has developed cells lines, Hs578T-ERαLuc and Hs578T-ERβLuc, with stable estrogen receptor elements fused to luciferase genes as well as separate inducible estrogen receptor (ER) constructs. Inducible ER expression will allow the use of high throughput screening for identifying ER-selective ligands inside cells.

Wis.L Master Cell Bank for Vaccine Development

UW–Madison researchers have developed a master cell bank of Wis.L cells (primary human fetal pulmonary fibroblasts) for commercial use as a substrate for the production of viral vaccines. The cells were first isolated by the State Laboratory of Hygiene (SLH) in 1968 and have been used extensively by SLH in viral diagnostic laboratories to isolate viruses from clinical specimens. The cells also have been used to grow rhinoviruses for inoculation studies. They have been subjected to testing as recommended by the FDA for use in vaccine production or experimental inoculation studies, and are free from extraneous contaminants.

Bone Morphogenetic Protein-1 Biological Materials

UW-Madison researchers have developed DNA clones, antibodies, vectors and cell lines for expressing bone morphogenetic protein-1 (BMP-1). These materials can be used to produce recombinant BMP-1.