Human bm mscs

Manufactured by Lonza

Sourced in Switzerland

Human BM-MSCs are mesenchymal stem cells derived from human bone marrow. They are an adherent, plastic-replicating cell type with the capacity for multi-lineage differentiation.

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Human MSCs (49 citations) BM-MSCs (34 citations)

12 protocols using human bm mscs

Isolation and Culture of Human Cell Lines

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Human BM-MSCs were obtained from Lonza (Basel, Switzerland) and were maintained with Mesenchymal Stem Cell Growth Medium (Lonza). The cells between passages 4 and 7 were used for all experiments. MDA-MB-231 cells and MCF7 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). MDA-MB-231 cells were cultured in Dulbecco’s modified Eagle medium/high glucose (DMEM/HG; HyClone, Logan, UT, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA) and 100 U/mL penicillin/streptomycin (P/S; Thermo Fisher Scientific). MCF7 cells were cultured in DMEM/F-12 (1:1) (Thermo Fisher Scientific) supplemented with 10% FBS, 2 mM L-glutamine (Thermo Fisher Scientific) and 100 U/mL P/S. Human umbilical vein endothelial cells (HUVECs; Lonza) were maintained on 0.2% gelatin from a bovine skin (Sigma-Aldrich, St. Louis, MO, USA)-coated dish using Endothelial Cell Growth Media-Plus (Lonza), and HUVECs between passages 4 and 8 were used for all experiments. All of the cells were maintained at 37 °C in a humidified incubator containing 5% CO₂.

Kim W., Park A., Jang H.H., Kim S.E, & Park K.S. (2022). Breast Tumor Cell-Stimulated Bone Marrow-Derived Mesenchymal Stem Cells Promote the Sprouting Capacity of Endothelial Cells by Promoting VEGF Expression, Mediated in Part through HIF-1α Increase. Cancers, 14(19), 4711.

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Bone Marrow Mesenchymal Stem Cell Encapsulation

Cited 2 times

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Human BM-MSCs used for in vitro viability experiments were obtained
from Lonza, Walkersville, MD. Cells were initially isolated from adult bone
marrow of bilateral iliac crests of normal, non-diabetic volunteers and
cryopreserved after passage 2. Passage 9 cells were encapsulated in
RLP-AM/HA-SH hydrogels at a density of 1 × 10⁶cells/mL.
In vivo experiments were completed using BM-MSCs with demonstrated
success in rabbit vocal fold scar [52 (link)]. Human BM-MSCs were obtained from Waisman Biomanufacturing,
Madison, WI. As previously described [52 (link), 53 (link)], cells were
derived from the iliac crest of a healthy 22-year-old female and expanded
and tested using procedures that met Current Good Manufacturing Practices
(cGMPs) for human clinical trials. Cells were suspended in cold freezing
medium containing PlasmaLyte, 10% rabbit serum albumin, and 2.5% dimethyl
sulfoxide (DMSO) at 2 × 10⁷ cells/mL and cryopreserved in
liquid nitrogen. Passage 6 cells were used in this study. Cells were thawed
just before injection, when they were gently mixed with RLP-AM/HA-SH gel via
pipetting at a final concentration of 3 × 10⁵ cells in 100
μL injection volume.

King R.E., Lau H.K., Zhang H., Sidhu I., Christensen M.B., Fowler E.W., Li L., Jia X., Kiick K.L, & Thibeault S.L. (2019). Biocompatibility and Viscoelastic Properties of Injectable Resilin-Like Polypeptide and Hyaluronan Hybrid Hydrogels in Rabbit Vocal Folds. Regenerative engineering and translational medicine, 5(4), 373-386.

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Human iPSC-MSCs and BM-MSCs Culture

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Human iPSC‐MSCs were derived from iPSCs (iMR90)‐4 (Lian et al. 2010 (link)). Human BM‐MSCs were purchased from the Lonza Walkersville, Inc. (Walkersville, MD). The cells were cultured as monolayers in DMEM (Thermo Scientific, Waltham, MA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA), 5 ng/mL bFGF (Invitrogen), and 5 ng/mL EGF (Peprotech, Hamburg, Germany).

Zhang J., Ho J.C., Chan Y., Lian Q., Siu C, & Tse H. (2014). Overexpression of myocardin induces partial transdifferentiation of human‐induced pluripotent stem cell‐derived mesenchymal stem cells into cardiomyocytes. Physiological Reports, 2(2), e00237.

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Bovine Meniscus and Human MSC Culture

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Normal bovine menisci were obtained from fresh bovine knees (Animal Technologies Inc.). To prepare meniscus explants, the avascular region was resected with a scalpel and cut into blocks of approximately 20-mm width. Cylindrical explants, 12-mm in diameter were harvested from the blocks by using a specimen needle and pre-cultured for 2–3 days at 37°C, 5% CO₂ until starting the experiment. Human BM-MSCs from 21– 23 years old donors were purchased from Lonza (Walkersville, MD) and cultured in Lonza MSCGM human Mesenchymal Stem Cell Growth BulletKit Medium (basal media for BM-MSCs).
Human meniscus and synovium were harvested from normal human knee joints. Vascular meniscus, avascular meniscus, and synovium were minced and digested separately with 2% collagenase for 12 hours. Synovial mesenchymal stem cells (SYN-MSCs) and meniscus cells were cultured in DMEM with 10% calf serum (CS), 1% penicillin-streptomycin-glutamine (PSG) (basal media for meniscus cells and SYN-MSCs).

Lee K.I., Gamini R., Olmer M., Ikuta Y., Hasei J., Baek J., Alvarez-Garcia O., Grogan S.P., D’Lima D.D., Asahara H., Su A.I, & Lotz M.K. (2020). Mohawk is a transcription factor that promotes meniscus cell phenotype and tissue repair and reduces osteoarthritis severity. Science translational medicine, 12(567), eaan7967.

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Culturing Human Bone Marrow-Derived Mesenchymal Stem Cells

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Human BM‐MSCs were purchased from Lonza (Walkersville, MD, USA) and cultured in 10% FBS α‐MEM under 5% CO₂ and 5% O₂ at 37°C.

Onodera Y., Teramura T., Takehara T., Obora K., Mori T, & Fukuda K. (2017). miR‐155 induces ROS generation through downregulation of antioxidation‐related genes in mesenchymal stem cells. Aging Cell, 16(6), 1369-1380.

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Magnetic Labeling of Bone Marrow Mesenchymal Stem Cells

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Human BMMSCs (Lonza, Basel, Switzerland) were cultured in MSCGM complete medium (Lonza, Basel, Switzerland) at 37°C under a 5% CO₂ atmosphere. Fresh medium was replaced every 3 days. BMMSCs between passages 4 to 10 were used in the experiments. For magnetic labeling 30 , 8 nm maghemite nanoparticles were used, synthesized by coprecipitation and stabilized in aqueous suspension by adsorption of citrate. These iron oxide nanoparticles (IONP) were dispersed in serum-free RPMI (Roswell Park Memorial Institute) medium (Sigma, France) supplemented with 5 mM citrate at a final iron concentration of 0.1 mM. BMMSCs underwent a 30-minute incubation with this medium at 37°C. Cells were then rinsed twice in serum-free RPMI medium. In order to allow nanoparticle internalization, cells were incubated for 2 hours with complete medium. Cells were fluorescently labeled with pKH67 green fluorescent dye, following the manufacturer's instructions (Sigma, St. Quentin Fallavier, France), and cultured in complete RPMI medium.

Rahmi G., Pidial L., Silva A.K., Blondiaux E., Meresse B., Gazeau F., Autret G., Balvay D., Cuenod C.A., Perretta S., Tavitian B., Wilhelm C., Cellier C, & Clément O. (2016). Designing 3D Mesenchymal Stem Cell Sheets Merging Magnetic and Fluorescent Features: When Cell Sheet Technology Meets Image-Guided Cell Therapy. Theranostics, 6(5), 739-751.

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Maintaining Undifferentiated H9-hESCs and Differentiating into M-MSCs

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Maintenance of undifferentiated H9-hESCs and differentiation into M-MSCs (Fig. 1a) were performed as previously described^{23 (link), 24 (link)}. The established M-MSCs were cultured with EGM2-MV medium (Lonza, San Diego, CA, USA) on plates coated with rat tail collagen type I (Sigma-Aldrich, St. Louis, MO, USA) in a humidified atmosphere with 5% CO₂ at 37 °C. All M-MSCs used in experiments were expanded less than ten passages to ensure multipotency. Characterization of basic features such as surface protein expression, cell proliferation, multipotency (in vitro differentiation into osteogenic, chondrogenic, or adipogenic lineages), in vitro angiogenesis assays, and karyotyping were performed as previously described^{23 (link), 24 (link)}. The M-MSC line stably expressing GFP was established by infection of GFP-expressing lentivirus produced as previously described^{14 (link)}. Human BM-MSCs purchased from Lonza (Basel, Switzerland) were cultured following the manufacturer’s instructions. Cells expanded for fewer than seven passages were used for experiments to ensure multipotency. All cells were tested for mycoplasm content each month (Mycoplasma Hoechst Stain Kit; 3030000; MP Biomedicals, LLC, Santa Ana, CA, USA).

Kim A., Yu H.Y., Lim J., Ryu C.M., Kim Y.H., Heo J., Han J.Y., Lee S., Bae Y.S., Kim J.Y., Bae D.J., Kim S.Y., Noh B.J., Hong K.S., Han J.Y., Lee S.W., Song M., Chung H.M., Kim J.K., Shin D.M, & Choo M.S. (2017). Improved efficacy and in vivo cellular properties of human embryonic stem cell derivative in a preclinical model of bladder pain syndrome. Scientific Reports, 7, 8872.

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BM-MSCs Culture and Migration Assay

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Human BM-MSCs were obtained from Lonza (Basel, Switzerland) and cultured in Mesenchymal Stem Cell Growth Medium (Lonza). TrypLE (Invitrogen, Carlsbad, CA, USA) was used for subculture, and the medium was changed every 3 days. For all experiments, BM‐MSCs between passages 5 and 7 were used. Dulbecco's Modified Eagle's medium (DMEM; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with heat-inactivated fetal bovine serum (FBS; Invitrogen), 1% L-glutamine (Invitrogen), and 1% penicillin and streptomycin (P/S; Invitrogen) was used for migration experiments using BM‐MSCs. MDA-MB-231 cells were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). MDA-MB-231 cells were cultured in DMEM/high glucose (DMEM/high, GE Healthcare Life Sciences), supplemented with 10% FBS and 1% P/S. MCF7 cells were obtained from the Korean Cell Line Bank (Seoul, South Korea). MCF7 cells were cultured in DMEM:Nutrient Mixture F-12 (DMEM/F-12; 1:1; Invitrogen), supplemented with 10% FBS and 1% P/S. All the cells were maintained at 37 °C in a humidified incubator containing 5% CO₂.

Choi S., Yu J., Kim W, & Park K.S. (2021). N-cadherin mediates the migration of bone marrow-derived mesenchymal stem cells toward breast tumor cells. Theranostics, 11(14), 6786-6799.

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Isolation and Expansion of Human Tendon, Bone Marrow, and Skin Cells

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Human tendon samples were obtained from semitendinosus tendon biopsies from 3 volunteers (3 men; mean age, 27 ± 1.4 years) undergoing anterior cruciate ligament reconstruction. The local ethics committee approved the study (PI2014108), and written informed consent was obtained from all donors. Tenocytes were isolated by explant culture as previously described.⁴ Human BM-MSCs (2 men, 1 woman; mean age, 32 ± 6 years) and skin fibroblasts (2 men, 1 woman; mean age, 39 ± 17 years) (not matched) were purchased from Lonza. The cells were cultured at 37°C in 5% CO₂, passaged at subconfluence, and used until passage 3; BM-MSCs were expanded in Dulbecco modified Eagle medium (DMEM) GlutaMAX (Gibco, Life Technologies) with 10% fetal bovine serum (FBS) (Hyclone; GE Healthcare), and skin fibroblasts and tenocytes were expanded in DMEM F12 supplemented with 10% FBS. Trypan blue was used to assess cell viability before plating.

Rubio-Azpeitia E., Sánchez P., Delgado D, & Andia I. (2017). Adult Cells Combined With Platelet-Rich Plasma for Tendon Healing: Cell Source Options. Orthopaedic Journal of Sports Medicine, 5(2), 2325967117690846.

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Osteogenic Differentiation of ST2 and BMMSCs

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ST2 cells were maintained in α-minimal essential medium containing 10% fetal bovine serum and 100 units/ml penicillin/streptomycin. Human BMMSCs were purchased from Lonza. Cell culture, and osteogenic differentiation were carried out following the instructions of the manufacturer (Lonza).

Zhou Y., Fyrner T., Chen C.H., Sather N.A., Hsu E.L., Stupp S.I, & Snead M.L. (2023). Optimization of peptide amphiphile-lipid raft interaction by changing peptide amphiphile lipophilicity. Acta biomaterialia, 164.

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