Early passage hAMSCs and BTIC cultures were used and authenticated by Johns Hopkins Genetic Resources Core Facility. hAMSCs (Invitrogen, R7788-115) were cultured in MesenPRO complete media (1% Antibiotic/Antimycotic (Invitrogen, 15240-062), 1% Glutamax (GIBCO, 35050-061), 1 vial of MesenPRO RS growth supplement (GIBCO, 12748-018), and MesenPRO RS basal media (GIBCO, 12747-010)). Human BTIC cultures (276 and 612) were obtained from intraoperative tissue (as approved by Johns Hopkins Institutional Review Board) and cultured in laminin-coated flasks (Sigma, L2020, 1 μg/cm2) with stem cell media (30 (link)). As previously validated and shown by our group, the human BTIC cultures are able to form oncospheres, are multipotential, and form tumors when implanted into animal models (30 (link)). To evaluate the tumorigenic capacity of BTICs in vivo, BTIC line 276 was injected intracranially in mice by our group resulting in the formation of solid tumors, while 612 formed diffuse tumors (30 (link), 31 (link)). The molecular subtype of BTIC culture 276 is mesenchymal and 612 is proneural, which was determined using a metagene score based approach for subtype designation, assessing four mesenchymal and two proneural genes using a microfluidics based qPCR assay (32 (link)). Commercial U87 cells (ATCC, HTB-14) were cultured in DMEM media with 10% FBS.
Mesenpro rs basal media
Manufactured by Thermo Fisher Scientific
Sourced in United States
MesenPRO RS basal media is a cell culture media designed for the maintenance and expansion of human mesenchymal stem cells (hMSCs). It provides a basal formulation optimized for the growth and proliferation of hMSCs.
Automatically generated - may contain errors
Lab products found in correlation
Mesenpro rs growth supplement, by Thermo Fisher Scientific (3 mentions)
Glutamax, by Thermo Fisher Scientific (3 mentions)
Collagenase a, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
L2020, by Merck Group (1 mentions)
Hamsc, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Moflo cytometer, by Beckman Coulter (1 mentions)
3 protocols using mesenpro rs basal media
1
Characterization of Brain Tumor Initiating Cells
2
Isolation and Transduction of hAMSCs
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Following approval by the Huazhong Science and Technology University Institutional Review Board, early passaged primary hAMSCs (hAMSC 173) were obtained from patients undergoing neurosurgical procedures as described in our previous studies14 (link),25 . The primary hAMSCs were isolated using the collagenase digestion method (collagenase-A; Thermo Fisher, Carlsbad, CA, USA) as described before13 (link). The cells were cultured in MSC complete media [MesenPRO RS basal media with one vial of MesenPRO RS growth supplement (Gibco), 1% Glutamax (Gibco), and 1% penicillin/streptomycin (Gibco)] and incubated at 37°C in a humidified atmosphere containing 5% CO2. Lentiviral vector–driven expression of VEGF (LV-VEGF-GFP) (Viraltherapy Technologies) was used to transduce the hAMSCs. VEGF expression was assessed by Western blot. All lentiviral (LV) constructs were packaged as LV vector in HEK 293 cells. After collection, the hAMSCs (hAMSC-Vector-GFP, hAMSC-Vector-GFP/Fluc, hAMSC-VEGF189-GFP, hAMSC-VEGF189-GFP/Fluc) were sorted by a Moflo cytometer (Beckman Coulter, Indianapolis, IN, USA).
3
Primary human adipose-derived mesenchymal stem cells isolation
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Following approval by the Institutional Review Board of Huazhong University of
Science and Technology, early passaged primary hAMSCs (TJH hAMSCs 019) were
obtained from patients during neurosurgical procedures, as described in our
previous studies20 (link),21 (link). The primary hAMSCs were isolated using the collagenase digestion method
(Collagenase-A, ThermoFisher, Carlsbad, CA, USA), following which they were
cultured in MSC media (MesenPRO RS basal media with one vial of MesenPRO RS
growth supplement [Gibco, Grand Island, NY, USA]; 1% penicillin/streptomycin
[Gibco]; and 1% Glutamax [Gibco]). Cells were incubated at 37 °C in a humidified
atmosphere containing 5% CO2.
Science and Technology, early passaged primary hAMSCs (TJH hAMSCs 019) were
obtained from patients during neurosurgical procedures, as described in our
previous studies20 (link),21 (link). The primary hAMSCs were isolated using the collagenase digestion method
(Collagenase-A, ThermoFisher, Carlsbad, CA, USA), following which they were
cultured in MSC media (MesenPRO RS basal media with one vial of MesenPRO RS
growth supplement [Gibco, Grand Island, NY, USA]; 1% penicillin/streptomycin
[Gibco]; and 1% Glutamax [Gibco]). Cells were incubated at 37 °C in a humidified
atmosphere containing 5% CO2.
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