GlobalStem is a provider of products and services for stem cell and neurobiology research

Transcription

1 GlobalStem is a provider of products and services for stem cell and neurobiology research 1

2 GlobalStem Products PluriQ Media Media, Sera and Buffers for Pluripotent Stem Cells 5 Feeder free Pluripotent Stem Cell Culture Media & Matrix 6 PluriQ Serum Replacement... 7 PluriQ Mouse Media..8 PluriQ Supplements Recombinant Mouse LIF (mlif). 9 FGF-2: Fibroblast Growth Factor-Basic. 9 GS21 Neural supplement Quality Control PluriPCR Kit.12 Stem Cell Reference Standard..13 Feeder Cell NuFF RQ ipsc Reprogramming Qualified Human Fibroblasts.14 Mouse Embryonic Fibroblasts (MEFs).15 Stem Cell BG01V Human Embryonic Stem Cells 17 GFP Expressing Mouse Embryonic Stem Cells 18 Primary and Neural Cell Products 19 HIP Neural Stem Cells.20 Primary Rat Cortical 22 Primary Rat CNS Stem Cells 23 2

3 Products for Stem Cell Research Feeder Cells and Stem Cell Reagents Mouse Embryonic (MEF ) & Human Newborn (NuFF) Fibroblasts Optimal support of healthy, undifferentiated ES & ips cell cultures Over 3 billion cells shipped every week. Consistent lot to lot performance & easy to use. Each lot is PluriQ TM tested on mouse and human ES cells. Contamination and pathogen free. NEW PluriQ TM Serum Replacement Defined serum free replacement for fetal bovine serum. Maintains healthy, undifferen ated human ES & ipsc cells in culture. Proven normal karyotype in hescs thru 20 passages. PluriQ TM bfbs, FGF, Conditioned Media, Freeze Media & and much more Primary and ips derived Neural Cells Primary Rat Cortical and Hippocampus Neurons HIP TM (Human Induced Pluripotent) Neural Stem Cells and Neurons Differentiates robustly to neurons. Capable of generating all the native cell types of the brain: neurons,astrocytes, andoligodendrocytes. Ready to go cryopreserved cells, just thaw and plate. 3

4 Media, Sera and Buffers for Pluripotent Stem Cells Ready For Action Media are made and tested by experts Concentratetion your research! Let GlobalStem help You with the tedious and meconsuming routine work of making and qualifying reagents. Ready For Action Media are optimized and tested for human and mouse embryonic stem (ES) cell and ips cultures. They are complete with non essential amino acids, β mercaptoethanol and dipeptide glutamine (a more stable approach to administering the necessary glutamine to ES/iPS cells). Combine Ready For Action Media with GlobalStem s Pluripotent Qualified Serum and Cryopreservation Media. Pluripotent Qualified Serum provides dependable growth and maintenance of undifferentiated human and mouse ES/iPSCs ensuring a consistent supply of cells for your experiments. Pluripotent Qualified Cryopreservation Media provide reliable post thaw viability for mouse and human pluripotent stem cells. Ready For Action PRODUCT CONTENTS CATALOG NUMBER PluriQ ES DMEM/F12 Optimized for human ES/iPS Cell culture 500 ml GSM 1002 PluriQ ES DMEM Optimized for mouse ES/iPS Cell culture 500 ml GSM 2001 PluriQ ES FBS Qualified for mouse and human ES/iPS Cell cultures 100 ml GSM 6002 Human hesfreeze Cryopreservation media 5 ml GSM 4200 Mouse mesfreeze Cryopreservation media 5 ml GSM 4100 GlobalStem products are for research use only. Extensive tesing guarantees dependable quality Starting with the finest media and sera, GlobalStem tests these reagents on multiple pluripotent stem cell lines to ensure that each batch provides healthy, dependable growth of embryonic stem and ips cells that are consistent, robustand enable undifferentiated proliferation. Each lot is also tested for contamination, including bacteria, fungi and mycoplasma, helping you avoid problems common to all cell culture work. 4

5 Feeder free Pluripotent Stem Cell Culture Media & Matrix Conditioned media for feeder free pluripotent stem cell culture GlobalStem offers Human Cell Conditioned Medium and Mouse Fibroblast Conditioned Medium that are complete ready to use, and performancetested for hes and ips cells. Just add basic fibroblast growth factor (bfgf). The Human Cell Conditioned Medium is serum free and conditioned with a proprietary human cell type. It ensures optimal culturing and pro liferation of undifferentiated human pluripotent stem cells consistent with feeder culture system. The Mouse Fibroblast Conditioned Medium is made using our high quality mouse embryonic fibroblast (MEF) cells. Human sourced extracellular matrix GlobalStem s Human Matrix is a proprietary, ready to use, human sourced extracellular matrix (ECM) free of human and mouse pathogens.just apply matrix to your plate, allow to coat overnight, aspirate and add human pluripotent stem cells. PRODUCT CONTENTS CATALOG NUMBER Pluripotent Serum Free, Feeder Free, Human 100 ml GSM 9200 Pluripotent Serum Free, Feeder Free MEF Conditioned Media 100 ml GSM 9100 Pluripotent Human Matrix 12.5 ml GSM 9250 GlobalStem products are for research use only. What our customers are saying... We started using GlobalStem Human Cell Conditioned Medium about six months ago. Our human ES cells are very robust, and remain healthy and undifferen ated for more than 10 passages. We are very pleased with the performance of GlobalStem Conditioned Medium for feeder free growth of our stem cells and highly recommend it to others. Luis Diaz, PhD / Johns Hopkins University We use GlobalStem Human Cell Conditioned Medium for culturing human ips cells. This rich medium has enabled us 5 to grow stem cells without serum or feeder cells. Our pluripotent cells appear healthy and robust for many passages. We tried alterna ves and believe this is an excellent product for feeder free growth of pluripotent cells. S. Gerecht,PhD / Johns Hopkins Univers

6 PluriQ Serum Replacement PluriQ Serum Replacement (PluriQ SR) is a defined, serum free supplement formulated as a direct replacement for FBS (fetal bovine serum) used in the growth and maintenance of undifferentiated human embryonic stem cells (hescs) and induced pluripotent stem cells (ipsc). PluriQ SR maintains healthy cell morphology and normal karyotype in hescs through 20 passages in culture. Maintenance of Pluripotency Phase Phase Oct4 Oct4 Figure 1. Oct4 Staining. Human ES cells cultured in PluriQ SR maintain pluripotency over extended passages. Images (left) show representa ve cell colonies stained with anti Oct4 after 11 passages Oct4 SSEA4 TRA-1-81 Figure 2. Pluripotency Markers. Flow Cytometry analysis of human ES cells co cultured in PluriQ SR for 20 passages show high expression of Oct 4 (left), SSEA4 (center), and TRA 1 81 (right). PRODUCT CONTENTS CATALOG NUMBER 500 ml GSM 6101 PluriQ Serum Replacement Supplement 100 ml GSM-6102 GlobalStem products are for Research Use Only. 6

7 PluriQ Serum Replacement Differentation Competence and Normal Karyotype after 20 Passages EB 1 week EB 2 weeks Figure 3. Human ESCs grown in PluriQ SR maintain normal karyotype for at least 20 passages. Figure 4. qpcr Data Analysis. After 20 passages, hescs cultured in PluriQ SR were able to form typical EBs (embryoid bodies) upon withdrawal of FGF. Using GlobalStem PluriQ PCR Kit for real time PCR analysis results show a decrease in gene expression and confirms the ability of cells to differentiat. Healthy, Undifferentiated Colony Morphology A B C Figure 5. Human ES cells cultured in PluriQ SR show normal, healthy colonies though 20 passages. Above images show representative colonies from passage 6 (A), passage 14 (B), and passage 20 (C). 7

8 Embryo Culture PrimeQ Mouse Embryo Culture Medium, with Phenol Red M2 medium supports the development of mouse embryos in culture and is used for collection handling of embryos outside the incubator for extended periods of time. M16 mouse embryo medium has been shown to support the development of mouse embryos during in vitro culture. PrimeQ KSOM Mouse Embryo Culture Medium,with AA & Phenol Red KSOM mouse embryo medium is commonly used for in vitro mouse embryo culturing KSOM and has been shown to support the development of mouse preimplantation embryos in culture. Additional studies have also shown that KSOM supplemented with amino acids increases the rate of blastocyst stage development.1,2,3 PrimeQ FHM Mouse Embryo Culture Medium,with Phenol Red FHM embryo culture medium is modification of KSOM where bicarbonate is partially replaced with HEPES buffer. FHM is typically used as an embryo collection media for washing and handling embryos outside the incubator. PRODUCT CONTENTS CATALOG NUMBER PrimeQ M2 Mouse Embryo Culture Medium (1X), with Phenol Red 50 ml GSM 5120 PrimeQ M16 Mouse Embryo Culture Medium (1X), with Phenol Red 50 ml GSM 5110 PrimeQ KSOM Mouse Embryo Culture Medium (1X), with AA & Phenol Red 50 ml GSM 5140 PrimeQ FHM Mouse Embryo Culture Medium (1X), with Phenol Red 50 ml GSM 5130 GlobalStem products are for Research Use Only. 8

9 Recombinant Mouse LIF (mlif) Leukemia Inhibitory Factor (LIF) is a lymphoid factor which is involved in a number of reproductive, inflammatory, developmental and regenerative processes. LIF promotes long term maintenance of pluripotent stem cells by suppressing spontaneous differentiation. LIF is also involved in a number of other biological processes including neural and hematopoietic cell differentiation, bone and fat metabolism, and mitogenesis of certain factor dependent cell lines. Each lot is PluriQ TM tested on mouse embryonic stem cells to confirm its functionality in plating efficiency, colony formation, morphology and maintenance of the undifferetintiated state. Applications Culturing of mouse pluripotent (ips/es) stem cells. Added as a supplement to cell culture medium to promote long term maintenance and propagation of undifferentiated mouse embryonic and induced pluripotent stem cells. Mouse Embryonic Stem Cells Cultured in GlobalStem s DMEM, 15% ES qualified FBS and 5 ng/ml (1000 U/ml) mlif. Max. 20X Phase Recombinant mouse LIF contains 188 amino acids and a 16 a.a. His dine based tag with a molecular mass of 22.8 kda. FGF-2: Fibroblast Growth Factor-Basic Human Recombinant FGF-2 Basic fibroblast growth factor is a member of the fibroblast growth factor (FGF) family. FGF family Members possess broad mitogenic and cell survival activities, and are involved in a variety of biological processes, including embryonic development, cell growth, morphogenesis, tissue repair, tumor growth and invasion. Human Recombinant Fibroblast Growth Factor-2 (FGF-2) is produced in E.coli. It is a single, non-glycosylated, polypeptide chain containing 155 amino acids and having a molecular mass of 17,353 Daltons. FGF-2 is purified by proprietary chromatographic techniques. 9

10 FGF-2: Fibroblast Growth Factor-Basic Applications Pluripotent (ips/es) cell culturing Human FGF-2 also promotes long-term maintenance and propagation of undifferentiated human embryonic and induced pluripotent stem cells. Culturing of neural stem cells human, rat & mouse FGF-2 is a necessary mitogen for the proliferation of neural stem cells and their maintenance of multipotency. Rat Neural Stem Cells Human ES Cells Neural stem cell marker: Nestin Alexa488 TM Nuclear marker: Dapi Phase Contrast Image PRODUCTS CONTENTS CATALOG NUMBER Leukemia Inhibitory Factor 100μg/vial GSR 7001 Fibroblast Growth Factor (FGF-2) 50μg/vial GSR-2001 GS21 Neural Supplement 10ml GSM 3100 GlobalStem products are for research use only. Double the testing to Ensure Quality Purity > 96%, by SDS-PAGE and RP-HPLC NIH-3T3 Fluorescence Activity Assay Activity Activity is measured using two cell proliferation assays, NIH-3T3 and BAF3 cells expressing FGF receptors, and The activity is >2 x 10 6 Units/mg. 10

11 GS21 Neural Supplement GS21 is a serum free neural media supplement specifically formulated to facilitate optimal health and long term viability in primary rat and mouse neurons in culture. It has been optimized from the published NS21 and B27 formulations 1,2. Improved Neuron Viability and Longevity Fig.1 Higher neuron viability after 2 & 4 weeks in culture Primary Rat Cortical Neurons (GSC 8220) cultured in GS21 supplemented culture media showed significantly higher neuron viability after 2 weeks, p=0.032 and 4 weeks, p=0.005 in culture, as compared to cells cultured in B27. Neurons plated in 96 well plates were labeled with MAP2 (green), and Dapi (blue) to denote nuclei. Data from n=4 wells is shown as mean ± SEM, 10X Mag. Fig.2 GS21Supplement supports 6 week cultured neurons Primary Rat Cortical Neurons cultured in GS21 supplementedculture media remain viable after 6 weeks in culture. Here, healthy, mature primary neurons iden fied by MAP2 staining (green), were imaged after 6 weeks in culture. Images were acquired at 10x and 20x (inset) magnification. 11

12 PluriPCR TM Kit Pluripotent Cell Line Assessment qpcr Kit GlobalStem s PluriPCR TM kit is designed as a quantitative, easy to use, and reliable assay of five genes strongly specific to pluripotency. These genes: Oct 3/4, Nanog, DNMT3b, Dppa4, and Rex1 are expressed by human embryonic stem cells (hesc) and induced pluripotent stem cells (ipsc); all are sharply downregulated during differentiation. Fully Optimized For Routine Quality Control Real time Quantitative PCR (qrt PCR) Developed for both the two step and one step qrt PCR methods Includes undifferentiated positive and differentiated negative control RNAs Robust normalization markers for accurate results Excel PCR Worksheet Simplified analysis and data interpretation Simplifying the Analysis and Data Interpretation GlobalStem has developed an assay that combines multiple pluripotency and housekeeping markers and an analysis tool to assess the pluripotency of your cell line. The PluriPCR Index is a simple method to understand if your pluripotent stem cells are differentiating. PluriPCR Index decreases as the pluripotent cells differentiate Amplification Plots (Ct Curves) Normalized Data PluriPCR Index PRODUCTS CONTENTS CATALOG NUMBER PluriPCR Kit 120 rxn Kit GSR 1001 GlobalStem products are for research use only. 12

13 Stem Cell Reference Standard Reference Standard of the International Stem Cell Initiative (ISCI) Embryonic Reference Standard Cells for Authentication and Characterization of Pluripotent Cells Flow Cytometry to Confirm Undifferentiaing Paraformaldehyde fixed, nullipotent human embryonal carcinoma line provides standard measurements for undifferentiated pluripotent cells (ips and ES). They are easily maintained as a monolayer in an undifferentiated, homogeneous population. Nearly 100% express cell surface and nuclear markers typical of undifferentiated hesc. The reference standard 2102Ep cells were Figure A: TRA 1 60 Alexa488 Figure B: SSEA 4 Alexa488 analyzed for multiple markers of pluripotency and differentiation using BD FACS Vantage SE/Diva flow cytometer. Two positive human ES markers (SSEA 4 and TRA 1 60) and one negative ES (SSEA1) marker were measured. Figures A and B represent the cell counts vs. the relative expression levels of TRA 1 60 and SSEA 4, respectively. Markers % Positive SSEA 4 97 TRA SSEA 1 2 Table 1 (left ) represents the % positive cells for ES markers SSEA 4, TRA 1 60 and SSEA 1 that confirm the cells are in an undifferentiatied state. The following an bodies were used: SSEA 4 (1:100, R&D Systems MAB1435); TRA 1 60 (1:100, Millipore MAB4360); and Alexa488 (1:750, Molecular Probes Invitrogen A11001). All lots are analyzed against representative pluripotent markers, including: SSEA3, SSEA4, TRA 1 60, TRA 1 81, and Oct4. PRODUCTS CONTENTS CATALOG NUMBER Flow Cytometry Reference Standard ESC cells Clone 4/D3 5 M cells/vial GSC 2002S Flow Cytometry Reference Standard ESC cells Clone 4/D3 2 M cells/vial GSC 2102S Gene Expression Reference Standard ESC cells 2/A6 5 M cells/vial GSC 2001 Gene Expression Reference Standard ESC cells 4/D3 5 M cells/vial GSC

14 NuFF RQ TM ipsc Reprogramming Qualified Human Fibroblasts During cellular reprogramming the feeder layer can greatly affect the health of the cell culture and the success of the reprogramming. While a range of mitotically inactivated fibroblasts can be used as a support layer for pluripotent cell culture, not all will support mrna reprogramming of patient dermal fibroblasts. GlobalStem offers Newborn Human Foreskin Fibroblasts (NuFF RQ ) that have been functionally validated by Stemgent TM to support the generation of induced pluripotent stem cell (ipsc) colonies using the Stemgent TM mrna Reprogramming Kit. Supporting Induced Pluripotent Stem Cell Reprogramming Features and Benefits: Pre Validated for ipsc Reprogramming provides optimal feeder layer support during mrna reprogramming. Pre Inactivated and Ready to Use Irradiated feeders facilitate faster and high efficiency primary ips cell generation. High Quality, Low Passage PluriQ TM tested fibroblasts provide consistent, long term support of healthy induced pluripotent stem cell cultures. Plate NuFF RQ TM feeder layer Plate target cells for reprogramming Transfect target cell NuFF RQ TM have been PluriQ TM tested and pre-validated by Stem-gent for support of healthy stem cell colonies during ipsc reprogramming. Pick primary ips Cells PRODUCT DESCRIPTION CONTENT CATALOG NUMBER NuFF RQ Reprogramming Qualified Human Fibroblast 4 5M cells/ vial GSC 3006G 14

15 Mouse Embryonic Fibroblasts (MEFs) Reproducibly grow undifferentiated pluripotent stem cell cultures Save time and trouble GlobalStem is the leader in MEFs for mouse and human pluripotent stem cells (ES and ips). Our MEFs are meticulously derived and comprehensively tested on mouse and human ES stem cells to ensure robust and consistent performance with every lot. Consistent performance GlobalStem specializes in pluripotent stem cell products and services. Quality and consistency is our trademark. Economical work low Save time and trouble dealing with an animal facility, performing tedious and time consuming dissections and siting at a hood to derive and expand cells. Safe handling Work with confidence. Comprehensive tesinng of each lot minimizes contamination risks to your lab. What our customers are saying... Ever since we stopped spending time making mouse embryonic fibroblasts and started getting them from GlobalStem we not only save a lot of time, but we also stopped worrying about mycoplasma and other contamination. Furthermore, GlobalStem MEFs are quite consistent lot to lot, which is very important. K. Halverson Transgenic Mouse Facility Institute of Neuroscience University of Oregon "Our lab has derived many batches of mouse embryonic fibroblasts (MEFs) and we have purchased MEFs from commercial sources. When our batches crashed, we were wary about purchasing MEFs, but decided to give GlobalStem a try. By comparison to other commercial MEFs, the GlobalStem MEFs are of a higher quality, very reliable and are more consistent from lot to lot. Our lab has been using GlobalStem MEFs for over two and half years and we are very satisfied." J. Miller Harvard Stem Cell Institute's hesc Core Facility 15

16 Mouse Embryonic Fibroblasts (MEFs) PRODUCT CONTENTS* CATALOG NUMBER CF 1 (Mitomycin C Treated) 7 8 M cells/vial High Density GSC 6101M CF 1 (Mitomycin C Treated) 4 5 M cells/vial Standard Density GSC 6001M CF 1 (Mitomycin C Treated) 2 M cells/vial Low Density GSC 6201M CF 1 (Mitomycin C Treated) 0.5 M cells/vial Ultra Low Density GSC 6301M CF 1 (Irradiated) 4 5 M cells/vial Standard Density GSC 6001G CF 1 (Irradiated) 2 M cells/vial Low Density GSC 6201G CF 1 (Irradiated) 0.5 M cells/vial Ultra Low Density GSC 6301G CF 1 (Untreated) 4 5 M cells/vial Standard Density GSC 6001 CF6Neo (Mitomycin C Treated, Neomycin Resistant) 7 8 M cells/vial High Density GSC 6105M CF6Neo (Mitomycin C Treated, Neomycin Resistant) 4 5 M cells/vial Standard Density GSC 6005M CF6Neo (Irradiated, Neomycin Resistant) 4 5 M cells/vial Standard Density GSC 6005G CF6Neo (Untreated, Neomycin Resistant) 4 5 M cells/vial Standard Density GSC 6005 DR4 (Irradiated, Neomycin, Hygromycin, Puromycin, 6 Thioguanine Resistant) DR4 (Irradiated, Neomycin, Hygromycin, Puromycin, 6 Thioguanine Resistant) 4 5 M cells/vial Standard Density GSC 6004G 2 M cells/vial Low Density GSC 6204G C57BL/6 (Mitomycin C Treated) 4 5 M cells/vial Standard Density GSC 6002M C57BL/6 (Irradiated) 4 5 M cells/vial Standard Density GSC 6002G C57BL/6 (Untreated) 4 5 M cells/vial Standard Density GSC 6002 C57BL/6 (Irradiated) 2 M cells /vial Low Density GSC 6202G *GlobalStem MEFs are provided at passage number 3. All GlobalStem products are for research use only 16

17 BG01V Human Embryonic Stem Cells A human ES cell line that is both easy to recover and culture BG01V human embryonic stem cells (hesc) are robust and easy to grow in cell culture. They remain undifferentiated for at least 25 passages. The fast growing and pluripotent BG01V hes cells recover beetter than most human ES cells and are ideal for use when getting started in human stem cell culturing. BG01V cells are derived from the wild type BG01 hes cells and are highly characterized. They are trisomic for chromosomes 12, 17 and XXY. Colonies of BG01V grown on irradiated or Mitomycin C treated mouse embryonic fibroblasts (MEFs) exhibit a predictable growth rate, and form large, uniform colonies. Colony of BG01V cells growing on mito cally arrested CF 1 MEF (GSC 6001M). Colony morphology is characteristic of undifferentiated hes cells. The pluripotency of the BG01V line has been demonstrated via teratoma formation [1]. This cell line responds to directed differentiation protocols in a manner similar to normal human ES cell line [2,3]. PRODUCTS CONTENTS CATALOG NUMBER BG01V Human Embryonic Stem Cell Line 1 vial GSC 1103 References Please note! BG01V is available for sale to non profit organizations only. Our Mouse Embryonic Feeder cells are ideal for use with the BG01V cells 1. Plaia TW et al Characteriza on of a new NIH registered variant human embryonic stem cell line, BG01V: a tool for human embryonic stem cell research. Stem Cells. 24(3): Zeng X et al BG01V: a variant human embryonic stem cell line which exhibits rapid growth after passaging and reliable dopaminergic differentiation. Restor Neurol Neurosci. 22(6): Pal R et all Similar pattern in cardiac differ entiation of human embryonic stem cell lines, BG01V and ReliCellhES1, under low serum concentration supplemented with bone morphogenetic protein 2. Differentiation. 75(2):

18 GFP Expressing Mouse Embryonic Stem Cells Engineering transgenic mice just got much easier Enhance your research with one or both of two new fullycharacterized, low passage, GFP expressing mouse embryonic stem cells (mescs) distributed exclusively by GlobalStem. LB10 mescs derived from a C57BI/6 mouse LC3 mescs derived from an F1 hybrid mouse (C57BI/6 x 129) Optimal GFP Expression Both mouse embryonic cells lines ubiquitously express Green fluorescent protein (GFP) driven by a CMV enhancer Chicken β actin promoter. Investigators can easily track GFP expressing cells in Transplantation experiments or in chimeric animals; the background genome is completely sequenced and annotated and experiments have shown the lines to be germ line competent. Superior Performance GlobalStem s experienced scientific team has expanded, characterized and cryopreserved (banked) both lines so that you can depend on their quality and performance for many passages. GlobalStem mescs are provided at low passage (P9) and express markers of the undifferentiated state. The C57BL/6 mouse has been chosen by the NIH Knockout Mouse Project (KOMP) as the preferred background for making null mutations in every gene in the mouse genome. The C57BL/6 mouse is the most widely used mouse strain and its genome has been completely sequenced and annotated. Live culture, GFP, B10 cells. Phase contrast image, live culture, LB10 cells. Fixed LB10 cells; Oct3/4 detection using rhodamine. PRODUCTS CONTENTS CATALOG NUMBER LB10 GFP Expressing Mouse ES Cells P9, 3 M cells/vial GSC 5003 LC3 GFP Expressing Mouse ES Cells P9, 3 M cells/vial GSC

19 Primary and Neural Cell Products PRODUCT Rat Primary Cortical Neurons Mouse Primary Cortical Neurons Rat Primary Astrocytes DESCRIPTION E18 Sprague-Dawley; Post-thaw viability typically 50%-75% Cells from other strains & brain regions available Post-thaw viability typically 50%-75% Cells from other brain regions available E18 Sprague-Dawley; Post-thaw viability typically >75% Cells from other strains & brain regions available Rat Primary Glial Restricted Precursors NeuroMax GS21 Supplement E18 Sprague-Dawley; Post-thaw viability typically >75% Cells from other strains & brain regions available Optimized to support long-term growth and viability in primary neurons in culture. Human Primary and ipsc-derived Neural Products PRODUCT Human Primary Neural Stem Cells DESCRIPTION Derived from normal healthy donor cells Human Primary Astrocytes Derived from normal healthy donor cells; Post-thaw viability typically >75% Human Primary Glial Restricted Precursors Human ipsc-derived Neural Stem Cells ipsc-derived Neurons Derived from normal healthy donor cells; Post-thaw viability typically >75% Derived from zero-footprint ipsc Line Available through our online catalog βiii-tubulin Positive Greater than 90% β-iii positive neurons Currently seeking Field Testers Key Features and Benefits: Cryopreserved cells with high post thaw viability Human & Rat Primary Neural Cells Human ipsc-derived Neural Cells Custom products and formats available Large scale cell manufacturing expertise 19

20 HIP Neural Stem Cells Pure Population of Human ipsc Derived Neural Stem Cells (NSCs) Introducing human ipsc derived NSCs for stem cell and CNS research. The NSCs are derived in consistent lots with confirmed normal karyotype. These NSCs were derived using a proprietary methodology and are predisposed to spontaneously differentiate into neurons. They also can be directed into astrocytes and oligodendrocytes. HIP NSC are derived from human induced pluripotent stem cells. These cells proliferate extensively in culture in response to basic fibroblast growth factor (bfgf), yielding a virtually pure population of neural stem cells. Virtually all cells express two widely accepted neural stem cell markers: Nestin (red) is expressed in greater than 99%, and Sox 1 (green) in greater than 97%. HIP Neural Stem Cells are Multipotent NSC s are capable of generating all the native cell types of the brain: neurons, astrocytes, and oligodendrocytes. HIP NSCs can differentiate along multiple pathways simultaneously. The image shows a differentiated HIP Neural Stem Cell culture made up of both β-iii Tubulin-expressing neurons (green) and Glial Fibrillary Acidic Protein (GFAP)-expressing astrocytes (red). Blue denotes DAPI staining of nuclei. HIP Dopaminergic Neurons HIP NSCs can be directed to differentiate efficiently toward a single neuronal subtype, such as Tyrosine Hydroxylase (TH)-expressing Dopaminergic neurons. 20

21 HIP Neural HIP Neural Stem Cells differentiate robustly to neurons Under controlled conditions, HIP NSCs can produce high yields of desired neurons. Differentiated cultures may contain greater than 90% neurons identified by β-iii Tubulin. HIP Neural Stem Cells Exhibit Various Neuronal Phenotypes Neuronal differentiation of HIP NSCs produces a high yield of neurons of various transmitier phenotypes. These include GAD67-expressing GABAergic neurons, ChAT-expressing Cholinergic neurons, and Glutamine Synthetase (GLSyn)- expressing Glutamatergic neurons (arrows). Tools for Research Ready-to-go just thaw and plate the cells according to our protocol with very good proliferation potential. Researchers who want high quality reproducible performance without the need of complicated isolation from tissue or the need to derive from pluripotent stem cell line. PRODUCT CONTENTS CATALOG NUMBER HIP Neural Stem Cells 1M Cells/ Vial GSC 4301 HIP Neurons & HIP Dopaminergic neurons Please Inquire Coming soon 21

22 Primary Rat Cortical The cerebral cortex plays an important role in memory, higher thinking, perception and the understanding of language. Our cryopreserved Primary Rat Cortical Neurons (RCN) are isolated from early embryonic stage (E18) Sprague Dawley Rats and provided cryopreserved as primary cells. Each vial of rat cortical neurons contains at least 4 million viable cells, with post thaw viability consistently in the range of 50 80%. Maximum Post thaw Viability Figure1. High viability during long term culturing Rat cortical neurons (GSC 8220) remain viable during long term culturing. Above images show cells at 1 week (A), 2 weeks (B) and 3 weeks (C) in culture. Figure2. Healthy, Mature Cortical Neurons Post thaw Primary rat cortical neurons (GSC 8220) were isolated from Sprague Dawley (E18) rats. MAP 2 (green) staining shows that greater than 90% of the isolated cells are neurons. Dapi staining (blue) denotes cell nuclei. PRODUCT CONTENTS CATALOG NUMBER Primary Rat Cortical Neurons 4M Cells/ Vial GSC 8220 GlobalStem products are for research use only. 22

23 Primary Rat CNS Stem Cells Ready for Expansion or Differentiation at Passage 0 Carefully prepared by highly experienced scientists, these Rat Neural Stem Cells from Sprague Dawley E14 cortex are ready for use just open the vial and plate. Our Rat CNS Stem Cells are multipotent and delivered at passage number zero (P0) in a quantity of about two million viable cells per vial. After plating, the cells are ready for expansion or differentiation into neurons, astrocytes and oligodendrocytes. They can be transfected, transplanted or differentiated into electrically active neurons there s no limit. Differentiated GlobalStem Rat Neural Stem Cells, co stained for the neuronal marker, Map2 (red), and the astrocyte marker, GFAP (green). The nucleitiare stained with DAPI (blue). Undifferentiated GlobalStem Rat Neural Stem Cells (NSC) stained for the NSC marker, nestin (green). The nuclei are stained with DAPI (blue). Extensively tested yet easily affordable GlobalStem Rat Neural Stem Cells are prepared under aseptic conditions and each lot must pass a meticulous screening process for bacterial, fungal, and mycoplasma contamination. All lots are assayed for multipotency. PRODUCTS CONTENTS CATALOG NUMBER Rat Neural Stem Cells (NSC) 1 vial; 2 million cells/vial; fully tested and verified GSC 8010 GlobalStem products are for research use only. 23

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