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

                                                                   STUDY PROTOCOL

EVALUATION OF ANTI-CANCER POTENTIAL OF PLANT EXTRACT(S)/HERBAL FORMULATION(S) USING HUMAN CANCER CELL LINE ORTHOTOPIC 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 and injected surgically into the cell line specific organ.

·       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 with paclitaxel 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 progression was followed by in vivo optical imaging system.

·       Animal’s blood will be withdrawn via retro orbital plexus and serum will be stored until analyzed.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

 

4.0  END POINT PARAMETER(S):

·       Clinical Observation

·       Body weight change

·       Tumor images

·       Tumor metastasis

5.0  REFERENCE(S):

5.1 Selene Nunez-Cruz, Denise C. Connolly, and Nathalie Scholler. An Orthotopic Model of Serous Ovarian Cancer in Immunocompetent Mice for in vivo Tumor Imaging and Monitoring of Tumor Immune Responses. J Vis Exp. 2010; (45): 2146.

                                   END OF DOCUMENT

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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 XENOGRAFT 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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