Primary osteoblasts were harvested from the bilateral parietal bone of 5-day-old Sprague–Dawley (SD) rats by enzymatic digestion, and bone marrow stem cells (BMSCs) were collected from the bone marrow extract by the adherent culture method (Animal Resources Centre of Guangxi Medical University, Nanning, Guangxi, China). After being stripped clearly with sterile gauze and digested with 0.25% trypsin−ethylene diamine tetraacetic acid (EDTA) (Beijing Solarbio Science and Technology Co., Ltd., China), the bilateral parietal bones were cut into pieces and digested with 1 mg/mL collagenase type I (Gibco BRL Co., Ltd., Gaithersburg, Maryland, USA) for 3 h. Meanwhile, bone marrow was flushed out of the femoral bone with culture medium. After centrifugation, isolated cells were suspended in alpha-modified Eagle’s medium (α-MEM, Gibco BRL Co., Ltd.) supplemented with 10% (v/v) fetal bovine serum (FBS, Zhejiang Tianhang Biotechnology Co., Ltd., Huzhou, Zhejiang, China) and 1% (v/v) antibiotics and cultured in a 5% CO2 incubator (Thermo Scientific TM Forma Series II Water-Jacketed, Santa Ana, California, USA) at 37 °C, with the medium changed every other day. At 80‒90% confluence after about 7 days of culture, primary cells were prepared for subsequent experiments.
Forma series 2 water jacketed
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Forma Series II Water-Jacketed is a laboratory incubator designed to maintain a controlled temperature environment for various applications. It features a water-jacketed design to ensure temperature stability and uniformity within the incubation chamber.
Automatically generated - may contain errors
3 protocols using forma series 2 water jacketed
1
Isolation and Culture of Rat Osteoblasts and BMSCs
2
Isolation and Culture of Primary Osteoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary osteoblasts were harvested from the bilateral parietal bone of newborn Sprague–Dawley (SD) rats (Animal Resources Center of Guangxi Medical University, Nanning, Guangxi, China) using enzymatic digestion. After being stripped clearly with sterile gauze and digested with 0.25% trypsin-ethylene diamine tetraacetic acid (EDTA) (Beijing Solarbio Science and Technology), the bilateral parietal bones were cut into pieces and subsequently digested with 1 mg ml−1 collagenase type I (Gibco BRL) for 3 h. After centrifugation, the isolated cells were resuspended and subsequently cultured at 37 °C in a 5% CO2 incubator (Thermo Scientific TM Forma Series II Water-Jacketed, Santa Ana, CA, USA), and the medium was changed every other day.
3
Isolation of Rat Parietal Osteoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary osteoblasts were harvested from the bilateral parietal bone of 3 to 7 days' newborn Sprague Dawley rats by enzymatic digestion. After rats were put to death by cervical dislocation, the bilateral parietal bones were stripped clearly with sterile gauze in a sterile environment. After the connective tissue around the parietal bone was removed by 0.25% trypsin-ethylene diamine tetraacetic acid (EDTA) (Beijing Solarbio Science and Technology Co., Ltd., Beijing, China), the bone was cut into pieces about 1×1 mm in a sterile vial and then digested with 1 mg/mL collagenase type I (Gibco-BRL, Carlsbad, CA, USA) in serum-free alpha-modified Eagle's medium (α-MEM, Gibco-BRL) for 3 h. After centrifugation at 1000 rpm for 5 min, isolated osteoblasts were suspended in α-MEM containing 10% (v/v) fetal bovine serum (FBS, Gibco-BRL) and 1% (v/v) antibiotics (penicillin 100 U/mL, streptomycin 100 U/mL). Cultures were maintained in a 5%-CO2 incubator (Thermo Scientific™ Forma Series II Water-Jacketed, Santa Ana, CA, USA) at 37℃ with the culture medium changed every other day. At 80-90% confluence after about 7 days of culture, primary cells were used for subsequent experiments.
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!