3.4 Methods and Materials
97
3.4.12 Mice Voluntary Cage-Wheel Exercise
BALB/c mice (female; 6 weeks old) were obtained from Vital River Laboratory
Animal Technology Co. Ltd. of Beijing, People’s Republic of China and allowed an
acclimation period of 1 week at 22 ± 2 °C with a 12-hour light: dark cycle (lights
on at 8am, lights off at 8 pm). Subsequently, BALB/c mice were randomly divided
into 2 groups (3–4 mice per group) and were subcutaneously injected with PBS or
peptide (10 mg/kg). Voluntary running was performed by these two groups at the
start of exercise following reported protocol. A voluntary running system consisting
of six separated chambers (Chengdu TME Technology Co., Ltd, China) was used
in the animal performance study. During the training session, mice were placed on
the motorized rod (30 mm in diameter) in the chamber. The rotation speed gradually
increased from 0 to 100 rpm over the course of 100 s. The rotation speed was recorded
when the animal fell off from the rod. Each rotarod training session consisted of 7
trials and lasted around 7 minutes. Performance was measured as the average rotation
speed animals achieved during the training session. The two different groups were
all trained at the same five continuous time points (day 0, day 2, day 4, day 8, day 12,
day 16, and day 20). No significant differences were found between the two groups.
3.4.13 In Vivo Imaging
When the tumor reached an appropriate volume of 200–300 mm
3 , the mice were
injected with 100 uL of Cy3-labeled PhR peptide by intratumoral injection. After
injection, mice were anesthetized with isoflurane. The induction concentration was
5% isoflurane/1L O 2 , and the maintenance concentration was 2–3% isoflurane/1L O 2 .
Once the mice were properly anesthetized, they were imaged at indicated time points
to monitor the metabolization of PhR
Cy3 peptide in tumors using the IVIS LuminaII
small animal in vivo optical imaging system (Caliper). In this study, a scanning
wavelength ranging between 500 and 950 nm was used for in vivo imaging.
References
1. Frank NY et al (2010) The therapeutic promise of the cancer stem cell concept. J Clin Investig
120(1):41–50
2. Ni C, Huang J (2013) Dynamic regulation of cancer stem cells and clinical challenges. Clin
Transl Oncol 15(4):253–258
3. Marhold M et al (2015) HIF1α regulates mTOR signaling and viability of prostate cancer stem
cells. Mol Cancer Res 13(3):556–564
4. Li Y, Laterra J (2012) Cancer stem cells: distinct entities or dynamically regulated phenotypes?
Can Res 72(3):576–580
5. Chen K et al (2013) Understanding and targeting cancer stem cells: therapeutic implications
and challenges. Acta Pharmacol Sin 34(6):732–740
97
3.4.12 Mice Voluntary Cage-Wheel Exercise
BALB/c mice (female; 6 weeks old) were obtained from Vital River Laboratory
Animal Technology Co. Ltd. of Beijing, People’s Republic of China and allowed an
acclimation period of 1 week at 22 ± 2 °C with a 12-hour light: dark cycle (lights
on at 8am, lights off at 8 pm). Subsequently, BALB/c mice were randomly divided
into 2 groups (3–4 mice per group) and were subcutaneously injected with PBS or
peptide (10 mg/kg). Voluntary running was performed by these two groups at the
start of exercise following reported protocol. A voluntary running system consisting
of six separated chambers (Chengdu TME Technology Co., Ltd, China) was used
in the animal performance study. During the training session, mice were placed on
the motorized rod (30 mm in diameter) in the chamber. The rotation speed gradually
increased from 0 to 100 rpm over the course of 100 s. The rotation speed was recorded
when the animal fell off from the rod. Each rotarod training session consisted of 7
trials and lasted around 7 minutes. Performance was measured as the average rotation
speed animals achieved during the training session. The two different groups were
all trained at the same five continuous time points (day 0, day 2, day 4, day 8, day 12,
day 16, and day 20). No significant differences were found between the two groups.
3.4.13 In Vivo Imaging
When the tumor reached an appropriate volume of 200–300 mm
3 , the mice were
injected with 100 uL of Cy3-labeled PhR peptide by intratumoral injection. After
injection, mice were anesthetized with isoflurane. The induction concentration was
5% isoflurane/1L O 2 , and the maintenance concentration was 2–3% isoflurane/1L O 2 .
Once the mice were properly anesthetized, they were imaged at indicated time points
to monitor the metabolization of PhR
Cy3 peptide in tumors using the IVIS LuminaII
small animal in vivo optical imaging system (Caliper). In this study, a scanning
wavelength ranging between 500 and 950 nm was used for in vivo imaging.
References
1. Frank NY et al (2010) The therapeutic promise of the cancer stem cell concept. J Clin Investig
120(1):41–50
2. Ni C, Huang J (2013) Dynamic regulation of cancer stem cells and clinical challenges. Clin
Transl Oncol 15(4):253–258
3. Marhold M et al (2015) HIF1α regulates mTOR signaling and viability of prostate cancer stem
cells. Mol Cancer Res 13(3):556–564
4. Li Y, Laterra J (2012) Cancer stem cells: distinct entities or dynamically regulated phenotypes?
Can Res 72(3):576–580
5. Chen K et al (2013) Understanding and targeting cancer stem cells: therapeutic implications
and challenges. Acta Pharmacol Sin 34(6):732–740