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Fresh bone marrow aspirate

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Fresh bone marrow aspirate is a laboratory product that provides a sample of bone marrow. It is collected through a procedure known as bone marrow aspiration. The sample can be used for various medical and research applications that require access to bone marrow.

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Lab products found in correlation

Phase contrast light microscope, by Zeiss (2 mentions) Stempro osteogenesis differentiation kit, by Thermo Fisher Scientific (2 mentions) Culture plates, by BD (2 mentions) Mem non essential amino acids solution, by Thermo Fisher Scientific (2 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions) 175 cm2 flask, by Corning (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Fungizone, by Thermo Fisher Scientific (1 mentions)

5 protocols using fresh bone marrow aspirate

1

Osteogenic Differentiation of hMSCs

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Films for cell culture were sterilized using ethylene oxide for 16 h at 4°C [40 (link)] and stored aseptically until seeding. hMSCs were obtained and cultured as described previously [41 (link)]. hMSCs were isolated from fresh bone marrow aspirates (Lonza, Basel, Switzerland), cultured in Dulbecco’s Modified Eagle Medium (supplemented with 10% fetal bovine serum, 0.1 mM non-essential amino acids, 1 ng/mL bFGF, 1% antibiotic/antimycotic) and seeded at passage 2 [41 (link)]. Cells were seeded at a density of 5,000 cells per cm2 in hMSC medium for 3 h, allowing the cells to adhere to the surface. Then, the medium was changed to osteogenic medium StemPro Osteogenesis Differentiation Kit (Gibco, Life Technologies, Grand Island, NY, USA). All cell cultures were performed in an incubator maintained at 37°C and 5% CO2. Cell growth and shape were monitored using a phase-contrast light microscope (Carl Zeiss, Jena, Germany).
Martín-Moldes Z., López Barreiro D., Buehler M.J, & Kaplan D.L. (2020). Effect of the silica nanoparticle size on the osteoinduction of biomineralized silk-silica nanocomposites. Acta biomaterialia, 120, 203-212.
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2

hMSCs Osteogenesis Differentiation Protocol

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Human mesenchymal stem cells (hMSCs) were cultured as described previously45 (link). Briefly, hMSCs were isolated from fresh bone marrow aspirates (Lonza, Basel, Switzerland), cultured in Dulbecco’s Modified Eagle Medium (supplemented with 10% fetal bovine serum, 0.1 mM non-essential amino acids, 1 ng/mL bFGF, 1% antibiotic/antimycotic) and seeded at passage 2. Prior to seeding, hydrogels were extensively extracted with deionized water to remove residual reactants and then incubated in cell culture media for 4 h at 37°C and 5% CO2. Cells were seeded at a density of 5,000 cells per cm2 in hMSC medium for 3 h, allowing the cells to adhere to the surface. Then, the medium was replaced with osteogenic medium StemPro Osteogenesis Differentiation Kit (Gibco, Life Technologies, Grand Island, NY, USA). All cell cultures were performed in an incubator maintained at 37°C and 5% CO2. Cell growth and shape were monitored using a phase-contrast light microscope (Carl Zeiss, Jena, Germany).
Martín-Moldes Z., Spey Q., Bhatacharya T, & Kaplan D.L. (2022). Silk-elastin-like-protein/graphene-oxide composites for dynamic electronic biomaterials. Macromolecular bioscience, 22(8), e2200122.
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3

Isolation and Expansion of Human Mesenchymal Stem Cells

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hMSCs were isolated from fresh bone marrow aspirate (Lonza) as previously described.29 hMSCs were expanded in Dulbecco's modified Eagle medium (DMEM) supplemented with fetal bovine serum (FBS) (10%), MEM Non‐Essential Amino Acids Solution (1%) (Gibco), penicillin streptomycin (PenStrep) (100 units/ml), and basic fibroblast growth factor (bFGF) (2 ng/ml) (Gibco). After expansion, hMSC was seeded on culture plates (Falcon, Tewksbury, MA) with a seeding density of 3.0 × 103 cells/cm2. The hMSCs were cultured in DMEM supplemented with FBS (10%), MEM Non‐Essential Amino Acids Solution (1%), and PenStrep (100 units/ml). All hMSCs used for this study were passage 4.
Takeda Y.S., Wang M., Deng P, & Xu Q. (2016). Synthetic bioreducible lipid‐based nanoparticles for miRNA delivery to mesenchymal stem cells to induce neuronal differentiation. Bioengineering & Translational Medicine, 1(2), 160-167.
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4

Isolation and Culture of Human Mesenchymal Stem Cells with Exosomes

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hMSCs were isolated from fresh bone marrow aspirate (Lonza, Allendale, NJ) as previously described [24 (link)]. hMSCs were expanded in DMEM, 10% FBS, 1% MEM Non-Essential Amino Acids Solution (Gibco), 100 units/mL Pen Strep, and 2 ng/mL basic fibroblast growth factor (bFGF) (Gibco). After expansion, hMSCs were seeded on culture plates (Falcon, Tewksbury, MA) at a seeding density of 3.0 × 103 cells/cm2. The hMSCs were then cultured in DMEM, 10% FBS, 1% MEM Non-Essential Amino Acids Solution, 100 units/mL Pen Strep. Exosome (40 μg protein/mL medium) was added to the culture medium every day. The culture medium was changed every 3 days. All hMSCs used for this study were at passage 4.
Takeda Y.S, & Xu Q. (2015). Neuronal Differentiation of Human Mesenchymal Stem Cells Using Exosomes Derived from Differentiating Neuronal Cells. PLoS ONE, 10(8), e0135111.
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5

Isolation and Expansion of hMSCs

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Human mesenchymal stem cells (hMCSs) were isolated from fresh bone marrow aspirate (Lonza, Walkersville, MD, USA) obtained from a single healthy, non-smoking male donor. Whole bone marrow cell isolate was plated at a density of 200,000 cells/cm2 in 175 cm2 flasks (Corning, Corning NY, USA). Dulbecco’s modified eagle medium (DMEM) with 10% fetal bovine serum (FBS), 0.1 mM non-essential amino acids, 1 ng/mL, 100 U/ml penicillin, 100 mg/mL streptomycin, and 0.25 ng/mL fungizone (Gibco, Grand Island, NY, USA) was used for expansion. Flasks containing a total volume of 20 mL were cultured in a humidified incubator at 37°C, 5% CO 2, and low oxygen (5%), and rocked daily to keep hemopoietic cells in suspension and allow stromal cells to adhere to the flask. The adherent cells were allowed to reach 90% confluence at which point they were trypsinized, suspended in FBS containing 10% DMSO, frozen at −80°C, and later stored in liquid nitrogen. Prior to experimentation, cells were thawed, re-plated, and passaged one more time before use.
McNamara S.L., Rnjak-Kovacina J., Schmidt D., Lo T.J, & Kaplan D.L. (2014). Silk as a biocohesive sacrificial binder in the fabrication of hydroxyapatite load bearing scaffolds. Biomaterials, 35(25), 6941-6953.
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