Human mesenchymal stem cells (hMSCs) were acquired from the Bioresource Collection and Research Center (Hsinchu, Taiwan). Moreover, they were maintained at 37 °C in humidified incubators with 5% CO2/95% air in specific culture media based on the protocol of our previous study, as described below16 (link). Briefly, hMSCs were cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with l -glutamine (4 mmol/L), 10% fetal bovine serum, and 1% penicillin–streptomycin. The confluent cells were expanded until passage 3, using 0.05% trypsin–EDTA. The final concentration was adjusted to 1 × 104 cells/mL, and aliquoted into 24-well Petri dishes (NUNCLON; Roskilde, Denmark). On the same day, DMEM was mixed with β-TCP control or β-TCP plasma treated at a concentration of 1 g/10 mL. Twenty-four hours later, the medium was removed from each test well and substituted for the test media, consisting of the previously described DMEM + β-TCP control or β-TCP plasma treated. Same Dulbecco’s modified Eagle’s medium first described was used on control wells.
Hmscs
Manufactured by RIKEN BioResource Center
Sourced in Japan
The HMSCs (Human Mesenchymal Stem Cells) are a type of multipotent stem cells derived from human sources. They have the ability to differentiate into various cell types, including osteoblasts, chondrocytes, and adipocytes. The HMSCs are widely used for research purposes in the fields of regenerative medicine, tissue engineering, and cell-based therapies.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Insulin transferrin selenium a, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Albumax 1, by Thermo Fisher Scientific (1 mentions)
Hyqtase, by Thermo Fisher Scientific (1 mentions)
Mcdb 201 medium, by Merck Group (1 mentions)
Carbonyl cyanide 4 trifluoromethoxy phenylhydrazone fccp, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Platelet derived growth factor, by Thermo Fisher Scientific (1 mentions)
L ascorbic acid 2 phosphate, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Scm141, by Merck Group (1 mentions)
Hmscs, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Heparin, by STEMCELL (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
4 protocols using hmscs
1
Culturing Human Mesenchymal Stem Cells with β-TCP
2
Optimizing Human Mesenchymal Stem Cell Culture
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Human mesenchymal stem cells (hMSCs) were purchased from the Bioresource Collection and Research Center (BCRC, No. RM60596), Hsinchu, Taiwan. ATCC (PCS-500-010). Cells were grown in culture dishes containing 56% Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA) and 37% MCDB-201 medium (Sigma Aldrich, St Louis, MO, USA), supplemented with 2% fetal bovine serum (Thermo, Logan, UT, USA), 1× Insulin-Transferrin-Selenium-A (Invitrogen, Carlsbad, CA, USA), 0.5 mg/mL of AlbuMax I (Invitrogen, Carlsbad, CA, USA), 10 ng/mL of epidermal growth factor (PeproTech, Rocky Hill, NJ, USA), 1 ng/mL of platelet-derived growth factor (PeproTech, Rocky Hill, NJ, USA), 10 nM dexamethasone (Sigma Aldrich, St Louis, MO, USA), and 50 μM L-ascorbic acid-2-phosphate (Sigma Aldrich, St Louis, MO, USA) [29 (link)]. The cells were maintained at 37 °C in a 5% CO2 incubator. When cells reached 70–80% confluence, they were detached with HyQtase (Thermo-Fisher Scientific, Waltham, MA, USA), centrifuged, and resuspended for subculture or further experiments. Cells between Passages 6 and 10 were used in the experiments. Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) (Sigma Aldrich, St Louis, MO, USA) was dissolved in DMSO as a stock solution at 10 mM, further diluted in hMSC cultured medium, and then used at various concentrations of 0.1, 0.3, 1, 2, 3, 10, and 30 μM.
3
Isolation and Culture of Rat and Human MSCs
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We collected bone marrow from the SD and CAG‐EGFP‐transgenic SD rats as described previously 22 , 33 . The cells were harvested in Dulbecco's modified Eagle's medium (DMEM; Sigma–Aldrich, St. Louis, MO) containing 10% FBS (Sigma–Aldrich) or STK1 (KBDSTC101; DS Pharma Biomedical) for the primary culture and were passaged after reaching 80%‐90% confluence. The culture medium in the STK1 group was then substituted with STK2 (KBDSTC102; DS Pharma Biomedical). The cells established under STK2 and medium containing 10% FBS were designated as “SF‐MSCs” and “10%MSCs,” respectively. Human MSCs from bone marrow (hMSCs; Riken BRC, Ibaraki, Japan) and human MSCs derived from adipose tissue (AT‐hMSCs; DS Pharma Biomedical) were similarly cultured in STK2 or cultured in DMEM containing 10% FBS. In human platelet lysate (HPL) analyses, rat MSCs and hMSCs were cultured in DMEM containing 10% heat‐inactivated HPL (SCM141; Merck Millipore, Billerica, MA) with 2 U/l heparin (#07980; STEMCELL Technologies, Vancouver, Canada). To inactivate HPL, HPL was incubated at 56°C for 30 minutes prior to use. Cells within passage 5 were used in all experiments and counted by using a TC‐20 (Bio‐Rad, Hercules, CA).
4
Isolation and Characterization of Rat and Human Mesenchymal Stem Cells
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rMSCs were isolated from bone marrow of the SD rat femur and tibia. rMSCs were cultured in DMEM containing 10% FBS (Sigma-Aldrich) for the primary culture. MSCs were passaged four times before used for administration or in vitro experiments. These cells were confirmed as MSCs by promoting their differentiation into osteocytes and adipocytes with specific differentiation media38 (link). Furthermore, we confirmed that standard MSC surface markers CD44 and CD90 were positive in these cells by flow cytometry (Supplementary Fig. S3 ). hMSCs derived from human bone marrow and human THP-1 monocytes were purchased from Riken BRC (Ibaraki, Japan). HK-2 cells, a human proximal tubular cell line, were obtained from the American Type Culture Collection (Manassas, VA). These cells were cultured as described previously29 (link).
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