INNOVATIVE PRODUCTS FOR NEUROSCIENCE RESEARCH

Transcription

1 INNOVATIVE PRODUCTS FOR NEUROSCIENCE RESEARCH NEURAL CELLS & MEDIA STEM CELLS & CULTURE REAGENTS TRANSFECTION CUSTOM SERVICES

2 NEUROSCIENCE PRODUCTS CATALOG TABLE OF CONTENTS Neuroscience research demands the highest quality cell culture products and reagents. MTI-GlobalStem has over 10 years experience providing neural and stem cell products and services, including cells, media, cell culture and transfection reagents. Our expertise is in neural cell isolation and characterization, scaling up stem cell production, storage, and banking and distribution of human induced pluripotent neural stem cells and neurons. We offer a complete portfolio of products you can trust for consistent results for neural cell culture for neuroscience and drug discovery research. ABOUT US Overview Who we are Our core competencies What we do.. 3 NEURAL CELL CULTURE Neuronal media & supplements 4 Neural stem cell media & supplements HUMAN IPSC-DERIVED NEURAL CELLS Human ipsc-derived neural stem cells... 6 Human ipsc-derived neurons... 7 PRIMARY NEURAL CELLS Stem cell culture media. 8 Stem cell culture supplements & growth factors... 8 NEURAL CELL TRANSFECTION Transfection reagents ORDERING INFORMATION Ordering information

3 ABOUT US COMPANY OVERVIEW WHO WE ARE MTI-GlobalStem, is a leading supplier of cutting-edge tools and reagents for the life sciences. MTI-GlobalStem has an active research program that utilizes expertise in transfection, neuroscience and stem cell biology to develop and supply scientists with the necessary tools and reagents to help advance new discoveries and developments in the life sciences. Utilizing our intellectual property and knowhow, and a specialized team of highly skilled and experienced scientists, we are filling the gap with primary and ipsc-derived neural cells, stem cells and supporting feeder cells, optimized growth media and supplements, and an innovative platform of novel targeted reagents used to deliver nucleic acids into stem cells and neuronal cells with maximum efficiency. The formation of MTI-GlobalStem merged Molecular Transfer, Inc. s (MTI) groundbreaking technologies in the areas of transfection, transformation and gene delivery, with GlobalStem s expertise in stem cell and neurobiology. A new corporate identity was announced in June 2015, featuring the re-branding of and GlobalStem as MTI-GlobalStem. Learn CORE COMPETENCIES WHAT WE DO NEUROBIOLOGY Our neurobiology portfolio includes neural cells, media and supplements to support neurotoxicity, drug screening or gene expression studies. Our proprietary cryopreservation process provides for consistent post-thaw viability of our neural cells. Researchers can confidently standardize their neural cell culture protocols with our high quality basal media and supplements. For your large-scale or technically challenging projects we offer custom services and products for neural cell isolation and differentiation. Contact for more details. NOVEL TRANSFECTION TECHNOLOGY We have developed a screening library of over 1000 proprietary lipid-formulations, enabling efficient identification and optimization of cell line and application specific reagents. With over 25 years of developing transfection reagents our R&D team has developed the most efficient products for DNA and RNA applications. STEM CELL CULTURE We develop and manufacture high quality stem cell culture reagents, including feeder cells, media and supplements for optimal, consistent culturing of pluripotent stem cells. Our products are used by scientists in over 500 research laboratories worldwide. CELL CULTURE MANUFACTURING Utilizing our highly trained personnel and expertise in large-scale primary and stem cell production, we produce over three billion cells each week. We have established a robust infrastructure and standardized quality control procedures to manufacture high quality cell reagents and support custom cell culture projects for our customers. Utilizing our intellectual property and know-how, and a specialized team of highly skilled and experienced scientists, we are committed to providing the highest quality products to support advancements in neuroscience. 3

4 NEURAL CELL CULTURE NEURAL CELL CULTURE MEDIA AND SUPPLEMENTS FOR NEURONS & NEURAL STEM CELLS Our neural basal media and serum-free supplements are optimized for maximum growth and survival of primary and ipsc-derived neurons and neural stem cells in culture. Our widely used serum-free GS21 neural supplement, based on the formulation of NS21 Supplement, has shown to support better viability than competitor reagents and eliminates the need for co-culture with astrocytes. Our neural media products include: NeuralQ Basal Medium (1X) GS21 Neural Supplement (50X) NeuralX NSC Medium (1X) GS22 Neural Supplement GS21 Neural Supplement w/o AO (50X) GS21 Neural Supplement w/o Steroids (50X) N2 Supplement (100X) ACHIEVE MAXIMUM NEURON VIABILITY AND CELL GROWTH Compe tor GS21 GS21 Supplement Top Compe tor NEURALQ MEDIUM & GS21 SUPPLEMENT Maximum cell viability during short-term and long-term 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 top competitor supplement. 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. We used the sample products [NeuralQ Basal Medium and GS21 Neural Supplement] you sent, and we are really pleased with our cultures. ~ SHEELA KADAPAKKAM UNIVERSITY OF TEXAS HEALTH SCIENCES CENTER, SAN ANTONIO GS21 SUPPLEMENT GS21 Supplement supports 6 week cultured neurons Primary Rat Cortical Neurons cultured in GS21 supplemented culture media remain viable after 42 days in culture. Here, healthy, mature primary neurons identified by MAP2 staining (green), were imaged after the 6 weeks and show healthy maturing neurons.. Images were acquired at 10x and 20x (inset) magnification. 4 We had the opportunity to compare your neuronal medium with to our best. I am surprised (and happy) ~ DR. RANDALL COHRS RESEARCH PROFESSOR, NEUROLOGY - UNIVERSITY OF COLORODO, DENVER

5 MEDIA AND SUPPLEMENTS FOR NEURAL STEM CELLS When maximum growth and survival of proliferative neural cells such as human ipsc-derived neural stem cells or rat primary oligodendrocyte precursors is needed, look to our NeuralX Basal Medium. NeuralX is intended for use with GS22 Neural Supplement (GSM-3200). When combined to form a complete medium, NeuralX and GS22 demonstrate superior performance in supporting the growth and survival of proliferating neural stem cells in culture. Improves long-term viability of proliferative neural cells. Supports more efficient and higher yields of oligodendrocyte precursors Serum-free, animal-free for minimal risk of introducing adventitious agents into the cell. NEURALX NSC MEDIUM & GS22 SUPPLEMENT Health Neural Stem Cell Culture ( Images, left) - NeuralX Neural Stem Cell Medium and GS22 Neural Supplement support healthy growth of proliferating human ipsc-derived neural stem cells in culture over multiple passages. HIP Human ipsc-derived pluripotent neural stem cells (BC1 line) were maintained in NeuralX NSC Medium and GS22 supplement for 4 days before passaging and continuing culture for another 4 days. Optimal Differentiation into Healthy, Mature Oligodendrocytes (Image, right) - Rat primary oligodendrocyte precursors (GRPs; GSC-9430) exhibit excellent growth, survival and health when grown in NeuralX NSC Medium and GS22 Neural Supplement. After rat GRPs were grown for 3 passages in NeuralX NSC Medium with GS22 supplement, antibody labeling indicated the cells were still primarily undifferentiated GRPs, as indicated by labeling with A2B5 (green) and DAPI (blue). SUPPORTING HEALTHY NEURAL STEM CELLS IN CULTURE N2 Supplement is a chemically defined, animal-free, supplement used for expansion of undifferentiated neural stem and progenitor cells in culture. N2 Supplement can also be used for maintenance of post-mitotic primary neurons in culture. N2 Supplement is intended for use with DMEM/F12 medium. While N2 can be used to grow primary neurons, we recommend NeuralQ Basal Medium (GSM-9420) with GS21 Neural Supplement (GSM-3100) for this application. N2 SUPPLEMENT N2 Supplement supports healthy growth of proliferating rat primary neural stem cells in culture over multiple passages. Primary Rat Neural Stem Cells (GSC-8010) were cultured in DMEM/F12 with N2 supplement. After 3 passages, cells were co-labeled with Sox2 (green) and Nestin (red). 5

6 HUMAN ipsc-derived NEURAL CELLS HUMAN ipsc-derived PRODUCTS NEURONS & NEURAL STEM CELLS Our human induced pluripotent stem cell (ipsc) derived neural cell products are extensively tested for quality and lot-to-lot consistency, providing reliable reagents for neurotoxicity, drug screening and gene expression applications. HUMAN ipsc-derived NEURAL STEM CELLS DIFFERENTIATE INTO NEURONS HUMAN ipsc-derived NEURAL STEM CELLS A B C D E Human ipsc-derived (HIP ) Neural Stem Cells generate a diverse neuronal population - HIP Neural Stem Cells derived from human induced pluripotent stem cells are multipotent. These cells proliferate extensively in culture and produce a virtually pure population of neural stem cells. Shown here, nearly all cells express two widely accepted neural stem cell markers, (A) Nestin is expressed in greater than 99%, and Sox 1 in greater than 97%. Under controlled conditions HIP NSCs produce high yields of mature neurons with differentiated cultures containing greater than 90% Tub3-positive neurons (B), including GAD67-expressing GABAergeric neurons (C), ChATexpressing Cholinergic neurons (D), and Glutamine Synthetase (GLSyn)-expressing Glutamatergic neurons (arrows) (E). HUMAN ipsc-derived NEURONS A B C D E Tub3/ Nestin Map2/Synapsin GFAP/Nestin Map2/ChAT Map2/Vglut1 HIP Neurons begin to show (A) high yields of maturing bipolar Tub3-positive neurons with some remaining elongated nestin-positive progenitors when differentiated for 4 days with GS21 Neural Supplement (GSM-3100). After five weeks of differentiation, cultures show (B) an abundance of MAP2-positive neurons expressing the synaptic marker, Synapsin, (C) astrocytes marked by GFAP, and (D) ChAT-positive cholinergic neurons. 6

7 HUMAN ipsc-derived NEURONS FOR DRUG DISCOVERY Pluripotent Stem Cells Neural Stem Cells Mature Neurons Neurons for Drug Discovery HIP Neurons exhibit post-synaptic activity. A) Representative voltage-clamp recordings from MTI-GlobalStem HIP Neurons following initial onset of activity revealed that the frequency of spontaneous synaptic events varied from cell-to-cell in the same culture. (B) A vertical scatter plot demonstrating a >100-fold range in mepsc frequency. (C) Following onset of activity (45 DIV) the number of neurons exhibiting activity were compared to the total number of neurons patched to determine the percentage of neurons that were active in each cell line. A total of 57.17% of HIP Neurons displayed activity (n = 21). Data Courtesy of Kyle S. Hubbard and Patrick McNutt, US Army Medical Research Institute of Chemical Defense, MD HUMAN ipsc-derived DOPAMINERGIC NEURONS Human ipsc-derived dopaminergic precursors differentiate into mature DA neurons- Human dopaminergic precursor cells were differentiated for 12 days using the GS-DOPA Kit of cells, media and supplements. Above image shows neuronal marker Tuj-1 (neuronal type β III tubulin) (green), Dapi (blue) nuclei stain. The dopamine release test (left chart shows functional GS-DOPA neurons), a quantitation of stimulated Dopamine release from cells after 12 days of differentiation. 7

8 PRIMARY NEURAL CELLS PRIMARY NEURAL CELLS RODENT NEURONS, NEURAL STEM CELLS & GLIA Why isolate your own neural cells? MTI-GlobalStem offers a wide range of cryopreserved rodent neural cells. Our proprietary method for cryopreserving cells produces neurons and glial cells with superior post-thaw viability, offering a convenient tool for your neuroscience applications. Our rodent neural cell products include: Primary Rat Neural Stem Cells Primary Rat Cortical Neurons Primary Mouse Cortical Neurons Primary Rat Hippocampal Neurons Primary Mouse Hippocampal Neurons Primary Rat Forebrain Neurons Rat Glial Restricted Precursor Cells Primary Rat Astrocytes Custom Neural Cell Isolation HIGH POST-THAW VIABILITY, FUNCTIONALLY IDENTICAL TO FRESHLY ISOLATED NEURONS PRIMARY RAT CORTICAL NEURONS Maximum Long-Term Viability - Primary Rat cortical neurons (GSC-8220) remain viable during long- term culture. Above images show cells at 1 week (A), 2 weeks (B) and 3 weeks (C) in culture. Evoked Synaptic Release - Cryopreserved versus freshly isolated primary rat neurons were each plated at Data courtesy of Pascal Laeng (Galenea Corp., MA). 80,000 cells per well and grown for 3 weeks. All the cells in the well were stimulated simultaneously. MTI-GlobalStem (GS, red trace) cryopreserved neurons exhibit evoked synaptic release responses virtually identical to that of freshly isolated rat forebrain neurons (blue trace). PRIMARY RAT NEURAL STEM CELLS PRIMARY ASTROCYTES A B Primary rat neural stem cells differentiate into neurons, co-stained for the neuronal marker, Tuß3(A, green), and the astrocyte marker, GFAP (B, green). The nuclei in both images are stained with DAPI (blue). 8 Healthy, Mature Astrocytes Primary Rat Astrocytes isolated from E19 Sprague-Dawley Rats. The above image shows differentiated Primary Rat Astrocytes characterized by GFAP (red) staining. Cells nuclei are stained with Dapi (blue).

9 NEURAL TRANSFECTION NEURAL TRANSFECTION DNA AND MESSENGER RNA DELIVERY REAGENTS Developed by the lead members of the scientific team that invented and developed Lipofectamine and Lipofectamine 2000, our specialized transfection reagents are formulations of proprietary compounds that are chemically defined and of animal-free origin. Routinely tested on a wide range of common cell lines our transfection reagents require minimal optimization for superior gene-delivery and expression results. SUPERIOR TRANSFECTION EFFICIENCY, MAXIMUM CELL VIABILITY Transfectasaurus Rex DNA-IN NEURO TRANSFECTION REAGENT Primary RCN Superior Transfection Efficiency in Neurons - DNA-In Neuro was used to transfect plasmid DNA encoding GFP into 6-day cultured E18 primary rat cortical neurons. Transfections were performed in 24-well plates using µl of DNA-In Neuro Reagent. Above data shows DNA-In Neuro significantly outperforms Lipofectamine 2000 (LF2K) with a near 3-fold improvement in transfection efficiency after 24 hours (right graph). Duplicate wells were assayed 24 hours (not shown) and 48 hours after transfection. The product [DNA-In Neuro] worked better than my current reagent. ~ NEUROSCIENCE RESEARCHER, DNA-In NEURO BETA TESTER LOW AMOUNTS OF MESSENGER RNA ACHIEVE MAXIMUM mrna DELIVERY IN NEURONS mrna-in NEURO TRANSFECTION REAGENT Primary RCN 500ng mrna 250ng mrna 120ng mrna Maximum efficiency with high expression with low amounts of mrna - With mrna-in Neuro, maximum transfection efficiency is achieved with reducing amounts of mrna, from 500ng (high) to 125ng (low). After 48 hours. mrna-in Neuro shows high efficiency in primary cortical neurons with uniform GFP expression. A Primary RCN B Maximum mrna transfection in neurons - Primary rat cortical neurons were transfected with 500ng mrna using mrna-in Neuro (A) vs LF2K (B). Images show significantly high efficiency and expression in cell body and neurites with mrna-in Neuro transfection after 24hrs. 9

10 ORDERING INFORMATION IPSC-DERIVED NEURAL CELL PRODUCTS DESCRIPTION CATALOG NO. HIP Neural Stem Cells HIP Neurons Human induced pluripotent stem (ips) cell-derived neural stem cells (NSC). These cells are predisposed to spontaneously differentiate into neurons and can be directed into astrocytes and oligodendrocytes. Unit size: 1M viable cells A mixed population of early differentiated neuronal progenitor cells derived from an integration-free human induced pluripotent stem (ips) cell line. Product includes cells, pre-coat matrix, and GS21 Neural Supplement (GSM- 3100). Unit size: 5M viable cells GSC-4301 GSC-4312 Dopaminergic Neurons GS-DOPA Kit Human ipsc-derived dopaminergic neural precursors (DNP) in a complete kit. Useful for the study of Parkinson's Disease and other neurological disorders. Mature, functional neurons obtained in days. Kit includes cells, growth medium and supplements. Unit size: 1M viable cells GSK-4401 PRIMARY NEURAL CELLS PRODUCTS DESCRIPTION CATALOG NO. Rat Cortical Neurons Rat Glial Restricted Precursors Rat Astrocytes Rat Neural Stem Cells Rat Hippocampal Neurons Rat Forebrain Neurons Mouse Cortical Neurons Mouse Hippocampal Neurons Isolated from early embryonic stage (E18) Sprague-Dawley Rats and provided cryopreserved as primary cell suspension. Unit sizes: 2M & 4M viable cells Isolated from newborn Sprague-Dawley Rats and provided cryopreserved at passage 2. Unit size: 1M viable cells Isolated from early embryonic stage (E19) Sprague-Dawley Rats and provided cryopreserved as primary cell suspension. Unit size: 1M viable cells Isolated from Sprague-Dawley (E14) rat cortex and provided as cryopreserved at passage 0. Unit size: 2M viable cells Isolated from early embryonic stage (E18) Sprague-Dawley Rats and provided cryopreserved as primary cell suspension. Unit size: 1M viable cells Isolated from early embryonic stage (E18) Sprague-Dawley Rats and provided cryopreserved as primary cell suspension. Unit size: 2M viable cells Isolated from E17 C57BL/6 embryonic mouse brain and provided cryopreserved as primary cell suspension. Unit sizes: 1M & 4M viable cells Isolated from E17 C57BL/6 embryonic mouse brain and provided cryopreserved as primary cell suspension. Unit sizes: 1M viable cells GSC-8220 GSC-8240 GSC-8040 GSC-8030 GSC-8010 GSC-8260 GSC-8202 GSC-8320 GSC-8340 GSC

11 NEURAL MEDIA & SUPPLEMENTS PRODUCTS DESCRIPTION CATALOG NO. NeuralQ Basal Medium (1X) GS21 Neural Supplement (50X) GS21 Neural Supplement w/o Antioxidants (50X) GS21 Neural Supplement w/o Steroids (50X) NeuralQ Basal Medium (1X) is optimized for maximum growth and longterm viability of neural cells in culture. Unit Size: 500mL GS21 is a serum-free neural supplement based on the formulation of NS21 Supplement and designed to improve the overall performance of neurons in culture. Unit Size: 10mL GS21 Supplement minus Antioxidants (AO) is the complete GS21 Supplement without 5 antioxidants (vitamin E, vitamin E acetate, superoxide dismutase, catalase, and glutathione); Designed for studies of oxidative stress in neurons. Unit Size: 10mL A customized formulation of the complete GS21 serum-free neural supplement without corticosterone and progesterone ideal for studying neuron-immune cell interactions. Unit Size: 10mL GSM-9420 GSM-3100 GSM-3110 GSM-3120 NeuralX NSC Medium (1X) GS22 Neural Supplement (50X) N2 Supplement (100X) NeuralX Neural Stem Cell (NSC) Medium is formulated for maximum growth and survival of proliferating neural stem cells in culture. Intended for use with GS22 Neural Supplement (GSM-3200). Unit Size: 500mL GS22 is a serum-free neural supplement specifically optimized for the propagation and long term viability of proliferative neural cells Unit Size: 10mL A chemically defined, animal-free, supplement used for expansion of undifferentiated neural stem and progenitor cells in culture. Can also be used for maintenance of post-mitotic primary neurons in culture. Unit Size: 10mL GSM-3110 GSM-3200 GSM-3300 NEURAL TRANSFECTION REAGENTS PRODUCTS DESCRIPTION CATALOG NO. DNA-In Neuro Transfection Reagent mrna-in Neuro Transfection Reagent Formulated specifically for maximum DNA delivery and cell viability in neurons and neural progenitors. Simplified singletube protocol. Unit sizes: 0.1ml,1 ml, 5x1 ml,10x1ml, Bulk Formulated specifically for maximum mrna delivery in primary and ipsc-derived neurons. Unit sizes: 0.1ml,1 ml, 5x1 ml,10x1ml, Bulk GST-2100 series GST-2110 series DNA-In SY5Y Transfection Reagent Optimized on the SH-SY5Y neuroblastoma cell line to give maximum transfection efficiency with exceptionally low cytotoxicity. Unit sizes: 0.1ml,1 ml, 5x1 ml,10x1ml, Bulk GST-2150 series INFO@MTI-GLOBALSTEM.COM SALES@MTI-GLOBALSTEM.COM 11

12 200 PERRY PARKWAY, SUITE 1 GAITHERSBURG, MD PHONE TOLL-FREE PHONE FAX INFO@MTI-GLOBALSTEM.COM 2015 MTI-GLOBALSTEM. ALL RIGHTS RESERVED PRODUCTS FOR RESEARCH USE ONLY NOT FOR THERAPEUTIC OR DIAGNOSTIC USES OCTOBER 2015 V4.0