EVALUATION OF ANTI-CANCER POTENTIAL OF PLANT EXTRACT(S)/HERBAL FORMULATION(S) USING HUMAN CANCER CELL LINE XENOGRAFT MODEL IN IMMUNECOMPRAMISED ANIMALS

                                                                       STUDY PROTOCOL

EVALUATION OF ANTI-CANCER POTENTIAL OF PLANT EXTRACT(S)/HERBAL FORMULATION(S) USING HUMAN CANCER CELL LINE XENOGRAFT MODEL IN IMMUNECOMPRAMISED ANIMALS

1.0  TEST SYSTEM DETAILS:

Species             : Mus Musculus

Strain                : Athymic Nude/ SCID

Age                   : 6-10 weeks

Body Wight        : 20-25 g

Sex                   : Male or Female

No. of animals    : 8 /Group

 

2.0  ALLOCATION OF GROUPS:

 

Groups	Treatment	Dose; ROA	No. of Animals
G1	Normal Control	Normal saline or 0.25% Na-CMC	8
G2	Reference Drug- Paclitaxel	10 mpk; i.p.	8
G3	Plant Extract/Herbal Formulation-1	X mpk; p.o.	8
G4	Plant Extract/Herbal Formulation-1	XX mpk; p.o.	8
G5	Plant Extract/Herbal Formulation-2	X mpk; p.o.	8
G6	Plant Extract/Herbal Formulation-2	XX mpk; p.o.	8

 

Extra 12 animals will be taken considering the tumor take up rate

Cell lines: HCT116, PANC-1, MDA-MB231,

 

3.0 METHOD:

·       Animals shall be procured from the CPCSEA authorized vendor.

·       Animals shall be quarantined for 1 week as per the in house SOP.

·       Healthy animals will be selected, randomized based on body weight and divided into 6 different groups consisting of 8 animals each.

·       Cell Culture:

·       Human cancer cell line will be procured from American Type Culture Collection USA, NCCS Pune or from authorized vendors cultured from suitable media supplemented with FBS or FCS or as applicable.

·       The cells were incubated in a humidified 5% CO₂ atmosphere at 37°C and harvested during the logarithmic growth phase.

·       The 1-15X10⁶/100µL cells in phosphate-buffered saline (PBS) or serum free media will be mixed with matrigel (1:1 ratio) and injected subcutaneously in the flank region of mice.

·       Tumor size will be measured by the digital vernier caliper and tumor volume will be calculated as 1/2(Length × Width²).

·       All the animals having tumor volume 50-200 mm³ will be distributed into different groups.

·       Drug treatment will be given as follows:

·       Group G1 animal will be treated as normal control and treated with normal saline or Na- CMC.

·       Animals of group G2 will be treated intraperitoneally weekly twice at the dose of 10mpk.

·       Group G3, G4, G5 and G6 animals will be treated with plant extract(s)/herbal formulation at different dose levels.

·       Tumor size will be measured weekly thrice until 28 days of drug treatment and compared with control animals.

·       Animal’s blood will be withdrawn via retro orbital plexus and serum will be stored until analyzed.

·       All the animals will be humanely sacrificed at the experiment.

 

4.0  END POINT PARAMETER(S):

·       Clinical Observation

·       Body weight change

·       Tumor volume

·       Tumor Growth Inhibition

 

5.0  REFERENCE(S):

5.1 Yuqin Yao, Yongjun Zhou, Xiaolan Su, Lei Dai, Lin Yu, Hongxin Deng, Lantu Gou, Andjinliang Yang. Establishment and characterization of intraperitoneal xenograft models by co-injection of human tumor cells and extracellular matrix gel. Oncol lett. 2015 dec; 10(6): 3450–3456.

                                     END OF DOCUMENT

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1. List of All SOPs and Documents for In-vivo Laboratory

2. List of All SOPs and Documents for Study Protocol

3. List of Pharmacological Study Models with details about disease Inducing agent, Standard or Reference Drug and related parameters.

4.List of OECD Guidelines or Toxicological Studies

5.List of Pharmacopeial Test or Quality Control Test or Batch Testing Parameters (In-vivo)

6.EVALUATION OF ANTI-CANCER POTENTIAL OF PLANT EXTRACT(S)/HERBAL FORMULATION(S) USING HUMAN CANCER CELL LINE ORTHOTOPIC MODEL IN IMMUNECOMPRAMISED ANIMALS

7.EVALUATION OF ANTI-CANCER POTENTIAL OF PLANT EXTRACT(S)/HERBAL FORMULATION(S) USING HUMAN CANCER CELL LINE ORTHOTOPIC MODEL IN IMMUNECOMPRAMISED ANIMALS

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