Mesenchymal stem cells (MSC) derived from bone marrow are a well-characterized population of adult stem cells. MSC have the capability for renewal and differentiation into various lineages of mature cells that produce fat, cartilage, bone,tendons, and muscle. These properties, in combination with their developmental plasticity, have generated tremendous interest in regenerative medicine to replace damaged tissues. These findings have spurred the development of MSC-basedtherapies for treating wide range of non-skeletal diseases ^1,2 .

iXCells Biotechnologies offers mBM-MSC isolated from mouse bone marrow. Each vialcontains >0.5 million cells. These cells are expanded in Mesenchymal Stem Cell Medium(Cat# MD-0037) and then cryopreserved at passage 2. iXCells mBM-MSC characterized by immunofluorescence and flow cytometry arestrongly positive for MSC marker CD29 [Figure 1]. These cells can further be differentiated into adipocytes using Adipocyte Differentiation Medium (Cat# MD-0005) and into osteoblastsusing Osteogenic Differentiation Medium (Cat# MD-0006) [Figure 2 and Figure 3]. mBM-MSC are negative for mycoplasma, bacteria, yeast, and fungi and can be expanded for no more than 3 passages in iXCells Mesenchymal Stem Cell Medium.

Figure 1. (A) Phase contrast image of MMSC-bm (B) Immunofluorescence staining with antibody against CD29. (C) Flow cytometric analysis shows >90% CD29 positive cells.

Figure 2. MMSC-bm adipocyte differentiation (Day 14 post adipogenic induction).

Figure 3. MMSC-bm osteogenic differentiation (Day 27 post osteogenic induction). Alizarin Red S staining of osteoblasts. The extracellular calcium deposit was stained in bright orange-red color.

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Reference

[1] Huang S, Xu L, Sun Y, Wu T, Wang K, Li G. An improved protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. JOrthop Translat. 2014, 27;3(1):26-33.

[2] García-Gómez I, Elvira G, Zapata AG, Lamana ML, Ramírez M, Castro JG, Arranz MG, Vicente A, Bueren J, García-Olmo D. Mesenchymal stem cells:biological properties and clinical applications. Expert Opin Biol Ther. 2010,10(10):1453-68.

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Mouse pulmonary endothelial cells are widely used in vascular biology and lung cell biology studies such as pulmonary inflammation (Biotech), angiogenesis, vessel permeability, leukocyte/EC interaction, nitric oxide production, and mechanotransduction ^[1]. Endothelial dysfunction is the common molecular basis of multiple human diseases, such as atherosclerosis, diabetes, hypertension, and acute lung injury. Primary culture of ECs is an important tool to dissect the role of endothelial genes in endothelial dysfunction-associated disorders ^[2]. Mouse pulmonary endothelial cells has been successfully used in phenotypic, and genetic studies characterizing endothelial genes in human diseases ^[3].

iXCells Biotechnologies provides high quality mouse pulmonary endothelial cells (MPEC), which are isolated from peripheral tissues of pulmonary lobes of C57BL/6 or CD1 mouse lung. MPECs are cryopreserved at passage 2 and delivered frozen. Each vial contains >0.5million cells. MPEC are characterized by immunofluorescence with antibody specific to Tie-2. MPECs are negative for mycoplasma, bacteria, yeast, and fungi. MPECs are guaranteed to further expand no more than 2 additional passages in Endothelial Cell Growth Media under the condition suggested by iXCells Biotechnologies.

Figure 1. Mouse pulmonary endothelial cells (MPEC). (A) Phase contrast image of MPEC. (B) Immunofluorescence staining with antibody against Tie-2.

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References

[1] Peramaiyan Rajendran, Thamaraiselvan Rengarajan, Jayakumar Thangavel, Yutaka Nishigaki, Dhanapal Sakthisekaran, Gautam Sethi, Ikuo Nishigaki. (2013) “The vascular endothelium and human diseases.” Int J Biol Sci. 9(10): 1057-69.

[2] Hadi AR Hadi, Cornelia S Carr, and Jassim Al Suwaidi. (2005) “Endothelial Dysfunction: Cardiovascular Risk Factors, Therapy, and Outcome”. Vasc Health Risk Manag. 1(3): 183-198.

[3] Monica J. Justice, Paraminder Dhillon. (2016) “Using the mouse to model human disease: increasing validity and reproducibility”. Disease Models & Mechanisms. Editorial.

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A ganglion is a group of nerve cells forming a nerve center, especially one located outside the brain or spinal cord. Dorsal root ganglion is a group of sensory nerve cell bodies. They pass sensory information to neurons in the spinal cord so it can be analyzed by the brain. In anatomy and neurology, the dorsal root ganglion (or spinal ganglion) is a nodule on a dorsal root that contains cell bodies of neurons in afferent spinal nerves. Dorsal root ganglion cells are pseudounipolar cells. Pseudounipolar cells have 2 axons rather than an axon and dendrite. One axon extends centrally toward the spinal cord; the other axon extends toward the skin or muscle.. Cultured adult rat dorsal root ganglion (DRG) neurons can be used to study depolarization-induced Ca²⁺ mobilization and the effects of intracellular Ca²⁺ depletion on neurite outgrowth. DRGs are very useful in neurotoxicity assessment & other drug screening studies.

Rat Dorsal Root Ganglion Neurons (rDRGN) provided by iXCells Biotechnologies are derived from spinal cords of normal embryonic rat. >500,000 cells are cryopreserved upon isolation.Each lot was tested for proper morphology, Stain positive for β III-Tubulin (TUJ1), Negative for HIV, Hepatitis B, Hepatitis C, mycoplasma, bacteria, and fungi.

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Epidermal keratinocyte is the predominant cell type in the outermost layer of the skin – the epidermis, which serves as a critical barrier to separate and protect the inside of human body from outside environment and damage from pathogens, heat, UV radiation and water loss. Epidermal keratinocytes originate in the stratum basale, and they undergo gradual differentiation and migrate towards the surface of the epidermis until they reach the stratum corneum, where they form a tight layer of nucleus-free and highly keratinized squamous cells. This layer forms an effective barrier to prevent water loss and the entry of infectious agents. Keratinocytes are also known to produce various growth factors, cytokines, antimicrobial peptides, and complement factors. Therefore, keratinocytes are important for wound healing, inflammation, infection, skin microbiome and immune response.

iXCells Biotechnologies provides high quality primary Mouse Epidermal Keratinocytes-adult (MEK-a), which are isolated from adult mouse tail and cryopreserved at P0, with ≥0.5 million cells in each vial. MEK-a are characterized by phalloidin staining and are negative for mycoplasma, bacteria, yeast and fungi. MEK-a are not recommended for expanding or long-term cultures.

Figure 1. (A). Phase contrast image of primary Mouse Epidermal Keratinocytes – Adult (MEK-a). (B). MEK-a were differentiated in the presence of 0.2mM CaCl₂, and cells were stained with phalloidin (red) to visualize the formation of actin fiber-rich filopodial projections between adjacent cells during differentiation. Nuclei were counterstained with DAPI.

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References

[1] Raja, Sivamani K, Garcia MS, Isseroff RR. Front Biosci. 2007;12:2849-68. Wound re-epithelialization: modulating keratinocyte migration in wound healing.

[2] Proksch E1, Brandner JM, Jensen JM. Exp Dermatol. 2008 Dec;17(12):1063-72. The skin: an indispensable barrier.

[3] Pasparakis M, Haase I, Nestle FO. Nat Rev Immunol. 2014 May;14(5):289-301. Mechanisms regulating skin immunity and inflammation.

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Skeletal muscle contains both differentiated myofibers and stem cells, known as satellite cells. The satellite cells, comprising around 1% of the total muscle nuclei, are situated between the plasma membrane of the multinucleated muscle cells and the basal lamina that surrounds each myofiber. In adult muscle, satellite cells are quiescent but proliferate in response to muscle injury, producing myoblasts that can either form new satellite cells or fuse with one another or pre-existing multinucleated muscle cells to help repair the muscle. They are responsible for postnatal muscle growth, hypertrophy and regeneration of skeletal muscle ^[1]. When quiescent satellite cells are activated, they co-express the transcription factors Pax7 and myoD ^[2].

Mouse Skeletal Muscle Satellite Cells (MSkMSC) from iXCells are isolated from mouse muscle of the pectoral girdle. MSkMSC are cryopreserved at passage one and delivered frozen. Each vial contains >5 x 10^5 cells in 1 ml volume. MSkMSC are characterized by immunofluorescence with antibodies specific to myosin, actin and actinin. MSkMSC are negative for mycoplasma, bacteria, yeast and fungi. MSkMSC are guaranteed to further expand for 15 population doublings using the culture medium provided by iXCells.

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Myoblast is a type of progenitor cells that differentiates to give rise to muscle cells ^[1]. They are the main effector cells in the myogenesis by fusing with each other to injured myofibers leading to the formation of new myofibers or regenerations of skeletal muscle in adults ^[2]. Many steps of myogenesis can be recapitulated through in vitro differentiation of myoblasts into myotubes. Primary myoblasts have been suggested as the most physiologically relevant model for studying myogenesis in vitro ^[3].

MSkMM from iXCells Biotechnologies are derived from mouse skeletal satellite cells isolated from mouse gastrocnemius (GA) and tibialis anterior (TA) muscles of both hind limbs by culturing them on matrigel-coated plastic dishes (Figure 1). MSkMM are cryopreserved at passage one and delivered frozen. Each vial contains ≥5 x 10⁵ cells/vial. MSkMM are characterized by immunofluorescence with antibodies specific to MyoD and Desmin (Figure 2). MSkMM can be in vitro differentiated into myotube using Mouse Skeletal Muscle Myoblast Differentiation Medium (Cat#MD-0065). MSkMM are guaranteed to further expand two passages using the culture medium provided by iXCells. These cells are negative for mycoplasma, bacteria, yeast and fungi.

Figure 1. Mouse Skeletal Muscle Myoblasts (MSkMM) were recovered and seeded at 1X10⁴/cm² in Mouse Skeletal Muscle Myoblast Growth Medium (Cat# MD-0064). Phase contrast images were taken at the indicated time post recovery.

Figure 2. Mouse Skeletal Muscle Myoblasts (MSkMM) were fixed at DIV1 and ICC staining was performed using antibodies against MyoD and Desmin, separately.

Figure 3. Mouse Skeletal Muscle Myoblasts (MSkMM) were cultured in Mouse Skeletal Muscle Myoblast Growth Medium (Cat# MD-Figure 3. Mouse Skeletal Muscle Myoblasts (MSkMM) were cultured in Mouse Skeletal Muscle Myoblast Growth Medium (Cat# MD-0064) until the cells reach 70-80% confluency. The growth medium was replaced with Mouse Skeletal Muscle Myoblast DifferentiationMedium (Cat# MD-0065) for 48 hours. The phase contrast image was shown indicating the formation of myotubes.

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References

[1] R L Perry, M A Rudnick. Molecular mechanisms regulating myogenic determination and differentiation. Front Biosci. 2000, 5: D750-67

[2] Roong Zhao, Alistair J Watt, Michele A Battle, Jixuan Li, Benjamin J Bondow, Stephen A Duncan. Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. Dev Biol. 2008, 317(2): 614–619.

[3] Brendan Evano, Shahragim Tajbakhsh. Satellite cells and the muscle stem cell niche. Physiol Rev. 2013, 93(1):23-67.

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Peripheral Blood Mononuclear Cells (PBMCs) is any blood cell with a single nucleus ^[1]. These blood cells are a critical component in the immune system to fight pathogen. PBMCs consist of lymphocytes (T, B and NK cells), monocytes and dendritic cells ^[2]. PBMCs are widely used in research fields including immunology, infectious diseases and hematological malignancies etc.

iXCells Biotechnologies provides high quality Mouse Peripheral Blood Mononuclear Cells (mPBMCs), which are isolated from mouse peripheral blood and cryopreserved with 5 million cells in each vial.

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Peripheral Blood Mononuclear Cells (PBMCs) is any blood cell with a single nucleus [1]. PBMCs consist of lymphocytes (T, B and NK cells), monocytes and dendritic cells [2].   PBMCs are widely used in research fields including immunology, infectious diseases and hematological malignancies etc.

iXCells Biotechnologies provides high quality Rat Peripheral Blood Mononuclear Cells (rPBMC), which are isolated from Sprague Dawley rat peripheral blood and cryopreserved.

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Macrophages are a type of white blood cell differentiated from circulating bone marrow-derived monocytes. Macrophages are responsible for detecting, engulfing and digesting cellular debris, apoptotic cells, and invading pathogens in a process called phagocytosis. Macrophages can be identified by several specific cell surface proteins including CD11b, CD14, F4/80 (mice)/EMR1 (human), MAC-1/MAC-3, and CD68 by immunohistochemistry or flow cytometry analysis ^[1]. T Bone marrow-derived macrophages are suitable for numerous applications including phagocytosis, gene expression profiling, and Morphological examination of cytospins using histological stains (e.g., May-Grünwald-Giemsa staining), etc.^[2].

iXCells Biotechnologies provides Mouse Bone Marrow Derived Macrophage (mBMDM), which were differentiated in the presence of M-CSF using the bone marrow cells isolated from adult C57BL/6 mice (Figure 1). mBMDM are harvested at P0 and delivered freshly or frozen. For frozen cells, each vial contains ≥ 5 million cells in 1 mL volume. mBMDMs are characterized by flow cytometry analysis with >97% CD11b+ purity (Figure 2). mBMDMs are negative for mycoplasma, bacteria, yeast, and fungi. It is recommended to use Macrophage Culture Medium (Cat # MD-0097) for in vitro culturing of mBMDM. However, mBMDMs are not recommended for long-term cultures or further expansion.

Figure 1. Mouse Bone Marrow Derived Macrophage (mBMDM). (A) Phase contrast images. (B) The purity of the mBMDM was measured using CD11b+ antibody by flow cytometry analysis.

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References

[1]. Cline MJ and Sumner MA. Bone Marrow Macrophage Precursors. I. Some Functional Characteristics of the Early Cells of the Mouse Macrophage Series. Blood. 1972; 40: 62-69.

[2]. Wynn TA and Barron L. Macrophages: master regulators of inflammation and fibrosis. Semin Liver Dis. 2010; 30: 245-57.

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