DW-MSCs were cultured in StemPro™ MSC SFM XenoFree medium (Gibco BRL, Grand Island, NY, USA) supplemented with 1% L-glutamine (200 mM) and 1% penicillin-streptomycin. The culture medium was changed every 2–3 days and the cells were maintained under 5% CO2 at 37°C. A549 cells (CCL-185, human type II AECs; ATCC, Gaithersburg, MD, USA) were cultured in DMEM supplemented with 10% FBS (Gibco BRL), 100 U/ml penicillin, and 100 mg/ml streptomycin. A549 cells were treated with 0 and 400 μM CoCl2 (SHINYO Pure Chemical, Osaka Japan) for 24 h and co-cultured with or without DW-MSCs. For cell viability assay, western blot analysis, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, and the mitochondrial respiration assay, DW-MSCs and A549 cells were indirectly co-cultured for 24 h using 0.4 μm Transwell™ inserts. For mitochondrial transfer analysis, DW-MSCs and A549 cells were directly co-cultured for 24 h.
Stempro msc sfm xenofree medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
StemPro MSC SFM XenoFree medium is a serum-free and xeno-free medium designed for the culture of human mesenchymal stem cells (hMSCs). It is formulated to support the expansion and maintenance of hMSCs without the use of animal-derived components.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (4 mentions)
Ccl 185, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Cellstart substrate, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Sodium l ascorbate, by Merck Group (1 mentions)
Glutamax 1 cts, by Thermo Fisher Scientific (1 mentions)
β glycerophosphate disodium, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Bm mscs, by PromoCell (1 mentions)
Ad mscs, by PromoCell (1 mentions)
Cellstart cts substrate, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Bioreactor, by Terumo BCT (1 mentions)
T175 cm2 flasks, by Thermo Fisher Scientific (1 mentions)
8 protocols using stempro msc sfm xenofree medium
1
Mesenchymal Stem Cells Rescue A549 Cells from Hypoxia
2
Expansion of Cryopreserved Bone Marrow Stromal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryopreserved BMSCs were thawed and plated at a cell density of 3000 cells·cm−2, on T-75 flasks with low glucose (1 g·L−1) Dulbecco’s Modified Eagle Medium [DMEM, (Gibco, Thermo Fisher Scientific, New York, NY, USA) supplemented with 5% v/v human platelet lysate (hPL) UltraGRO™-PURE (AventaCell, Atlanta, GA, USA) and Antibiotic-Antimycotic (1×) (Gibco, Thermo Fisher Scientific) (DMEM/HPL). Alternatively, cells were plated at the same cell density in CELLstart™ substrate (Gibco, Thermo Fisher Scientific) pre-coated T-75 flasks with StemPro™ MSC SFM XenoFree medium (Gibco, Thermo Fisher Scientific) supplemented with Glutamax (1×) (Gibco, Thermo Fisher Scientific) and Antibiotic-Antimycotic (1%) (StemPro). Cells cultured in DMEM-HPL were grown in VWBR, whereas cells grown in SP were cultured in StemPro. At 70% cell confluence, MSCs were harvested with 1× TrypLE™ Select Enzyme solution (Gibco, Thermo Fisher Scientific) for 5 min at 37 °C. Cell number and viability were determined using the Trypan Blue (Gibco, Thermo Fisher Scientific) exclusion method.
3
Expansion and Characterization of Human AD-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human adipose tissue-derived MSCs (AD-MSCs; passage 2) were obtained from PromoCell (Heidelberg, Germany; catalog number C-12977) and expanded until passage 4 using StemPro MSC SFM XenoFree medium (Thermo Fisher Scientific, Waltham, MA, USA) in low oxygen (5% O2) in the presence of 5% CO2 at 37 °C. Cells were tested by PromoCell for morphology, proliferation potential, adherence rate, and viability. Cells were analyzed by flow cytometry using a comprehensive panel of markers, CD73/CD90/CD105 and CD14/CD19/CD34/CD45/HLA-DR. Adipogenic, osteogenic, and chrondrogenic differentiation assays were performed for each lot in the absence of antibiotics and antimycotics.
4
Osteogenic Differentiation of Adipose-derived MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adipose-derived MSCs were obtained from Lonza (Walkersville, MD, USA) and cultured in serum free StemPro® MSC SFM XenoFree medium (Thermo Fisher Scientific, Waltham, MA, USA) containing CTS™ GlutaMAX™-I (Thermo Fisher Scientific) at 37 °C in an atmosphere with 5% CO2. For osteogenic differentiation, passage 3–6 ADSCs were cultured to 90% confluence, and then the culture medium was replaced with osteogenic medium [MSC growth medium supplemented with 5 nM dexamethasone (Sigma-Aldrich, St. Louis, MO, USA), 250 μM (+)-sodium L-ascorbate (Sigma-Aldrich), and 10 mM β-glycerophosphate disodium (Sigma-Aldrich)]. The medium was changed two or three times a week, and the cells were cultured for 21 days.
5
Characterization of Human MSCs from Bone Marrow and Adipose Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human bone marrow-derived (BM-MSCs; passage 2) and adipose tissue-derived MSCs (AD-MSCs; passage 2) were obtained from PromoCell (Heidelberg, Germany; C-12977). The cells were expanded through passage 4 in StemPro MSC SFM XenoFree medium (Thermo Fisher Scientific, Waltham, MA, USA) under hypoxic (5 %) conditions in the presence of 5 % CO2 at 37 °C.
The morphology, proliferation potential, adherence rate, and viability of the cells were tested using PromoCell. The cells were analyzed by flow cytometry using a comprehensive panel of markers, CD73/CD90/CD105 and CD14/CD19/CD34/CD45/HLA-DR. Adipogenic, osteogenic, and chondrogenic differentiation assays were performed for each lot in the absence of antibiotics and antimycotics.
The morphology, proliferation potential, adherence rate, and viability of the cells were tested using PromoCell. The cells were analyzed by flow cytometry using a comprehensive panel of markers, CD73/CD90/CD105 and CD14/CD19/CD34/CD45/HLA-DR. Adipogenic, osteogenic, and chondrogenic differentiation assays were performed for each lot in the absence of antibiotics and antimycotics.
6
Isolation and Culture of hUC-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human umbilical cords (hUCs) were collected from West China Women’s and Children’s Hospital without any complications of pregnancy or parturition, and the collected hUCs were transferred in sterile boxes that contained cold Hanks’ Balanced Salt Solution (HBSS) (Life Technologies, USA). Written informed consent was obtained from the pregnant women before labor. This study was approved by the institutional ethics committee of Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC).
hUCs were dissected longitudinally, and the arteries and veins were removed. The remaining pieces were chopped into 0.2cm3 size. These explants were transferred in the CELLstart™ CTS™ Substrate (Life Technologies, USA)-coated 100 mm plates (Nunc, Denmark). Then, a few StemPro® MSC SFM XenoFree medium (Life Technologies, USA) with 1% penicillin-streptomycin (Life Technologies, USA) was added to the plates, and the explants were cultured at 37°C in a 5% CO2 incubator and left undisturbed to allow the cells to migrate from the explants. After 7–9 days, the MSC-like cells were found around the fragments. The cells were passaged into another plates and further split 1:4 by 0.05% Trypsin-EDTA (Life Technologies, USA) once the cells reached 80% confluence.
hUCs were dissected longitudinally, and the arteries and veins were removed. The remaining pieces were chopped into 0.2cm3 size. These explants were transferred in the CELLstart™ CTS™ Substrate (Life Technologies, USA)-coated 100 mm plates (Nunc, Denmark). Then, a few StemPro® MSC SFM XenoFree medium (Life Technologies, USA) with 1% penicillin-streptomycin (Life Technologies, USA) was added to the plates, and the explants were cultured at 37°C in a 5% CO2 incubator and left undisturbed to allow the cells to migrate from the explants. After 7–9 days, the MSC-like cells were found around the fragments. The cells were passaged into another plates and further split 1:4 by 0.05% Trypsin-EDTA (Life Technologies, USA) once the cells reached 80% confluence.
7
Bioreactor Culture of hUC-MSCs for Conditioned Medium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bioreactor (TERUMOBCT, Lakewood, CO, USA) was fed via two circulation loops. The internal loop and the extra capillary loop, both of which have inlets for media and reagents or cells (external loop only) [14 (link)]. The medium used to cultivate hUC-MSCs in the bioreactor was the same medium used for the hUC-MSCs in the FM. The hUC-MSC medium for bioreactor cultivation was obtained from Prime Cell Therapeutics Inc., Czech Republic. The culture method was automatically set by a program embedded in the bioreactor.
The hUC-MSCs were plated at 5 × 105 cells and were cultured in FBS-free DMEM (StemPro MSC SFM Xeno Free medium, Life Technologies, Gaithersburg, MD) for eight days.
The culture supernatant was collected in a bag attached to the bioreactor, filtered through a 0.2 mm filter, and stored at −80 °C. This conditional medium was labeled as BM to indicate that it was obtained following the systematic culture of hUC-MSCs in a bioreactor.
The hUC-MSCs were plated at 5 × 105 cells and were cultured in FBS-free DMEM (StemPro MSC SFM Xeno Free medium, Life Technologies, Gaithersburg, MD) for eight days.
The culture supernatant was collected in a bag attached to the bioreactor, filtered through a 0.2 mm filter, and stored at −80 °C. This conditional medium was labeled as BM to indicate that it was obtained following the systematic culture of hUC-MSCs in a bioreactor.
8
Expansion of Human Umbilical Cord Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hUC-MSCs were resuspended at 5 × 105 cells/cm2 in T-175 cm2 flasks (Thermo Fisher Scientific, USA), and hUC-MSCs were cultured in FBS-free DMEM medium (StemPro MSC SFM Xeno Free medium, Life Technologies, Gaithersburg, MD) for six days. Upon reaching 80% confluence, the monolayer was washed and harvested using Trypsin–EDTA (Thermo Fisher Scientific, USA), and the cells were split 1:4 into a new T-175 flask. The cells routinely replaced the medium when 80% of the mating was reached, which took approximately three days. The cultured medium was collected with a 0.2 μm filter replaced each time with the medium and stored in a deep freezer at −80 °C. We named it FM, which was kept in the flask.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!