Table 4 The implantation mouse models of hepatocellular carcinomas

McClendon et al. [81]

HCC

FVB

Alb-cre mice were crossed to conditional Rb and/or p53 KO mice to generate a model for tissue-specific inactivation of Rb and p53. 14-day old mice were given a single interperitoneal injection (20 mg/kg) of DEN.

Duchosal et al.[86]

The hu-PBL-SCID mouse model

SCID

6 week-old SCID mice were injected PBL isolated from diluted (1:2) blood.

Ito et al. [87]

NOD/scid/γ^null_c

mouse model

NOD/Shi-scid

C57BL/6J-γc null

Female NOD/Shi-scid mice were crossed with male C57BL/6J-γ cnull mice, F1 females were mated with NOD/Shi-SCID males. Males obtained were backcrossed 7 times with NOD/Shi-SCID mice. Mice obtained by 8 backcrossings were intercrossed to obtain mice homologous for the SCID and γ c null genes.

Traggiai et al. [90]

Cord blood cell–transplanted mice

Rag2-/-γc-/c-

We transplanted newborn Rag2-/- γc-/c- mice CD34 cord blood cells. Mice were subsequently analyzed between weeks 4 and 26 of age, until human CD45 hematopoietic cells were detected in all animals.

Khodayari et al. [91]

implantation mouse models

BALB/c

After were radiated by Cobalt-60 (4 Gy) 24 h the female BALB/c mice (6–8 weeks) have subcutaneously received 3 × 10⁶ MCF-7 cells in the right flank.

Yao et al.[94]

implantation mouse models

BALB/c

1. HepG2 transplantation tumor model: HepG2 cells (2 × 106) were subcutaneously injected into each mouse. Tumor-bearing mice were grouped according to the tumor volume after one week.

2. PDX model: All fragments from one hepatoma patient were subcutaneously inoculated into one flank of the experimental 5-week-old nude mice. Tumor growth was measured twice weekly using a Vernier caliper. The established PDX model was called passage 1 (P1). When the tumor size of P1 reached approximately 750 mm ³, the tumor was separated and sliced into small fragments (approximately 3 × 3 × 3 mm³/fragment) and reinoculated into mice to obtain the subsequent passages P2, P3, P4, and so on.

Su et al.[95]

Caudal vein injection mouse model and implantation mouse models

BALB/c

NOD/SCID

1. HepG2 and MHCC97H cells with/without YTHDF1 knockdown were exposed to sublethal heat treatment and recovered for 1 h before injected into the tail vein of NOD/SCID mice.

2. Subcutaneous tumors were first grown through inoculating HCCLM3-NC and HCCLM3-shSTIP1 cells (1 × 107 cells/ spot) at the right flank of mice. When reached 10-mm in diameter, tumors were harvested, non-necrotic tissues were cut into 1 mm³ pieces and implanted into the left lobe of another tumor-free mouse’s liver. The experiment was carried out four weeks after implantation.

3. After washing out of blood and unwanted tissues, the tumor blocks were cut into pieces at 1 mm × 1 mm × 1 mm under sterilized condition. Mice were anesthetized. And a 1 cm subcutaneous pocket was made on the right flank to store the tumor piece. About three months later, successful subcutaneous xenograft was visible and could be stably passed from one mouse to another. We anesthetized the tumor-bearing mice again to harvest the tumors and cut them into pieces at 1 mm×1 mm×1 mm again. Another batch of mice was anesthetized. Subcostal incision was performed to expose liver lobes. Tumor pieces were placed into the liver via a tunnel made by microscopic forceps. Mice were resumed feeding to nourish orthotopic tumor. Successful orthotopic tumor could be palpated within 2 months.

Xun et al. [96]

implantation mouse models

NOD/SCID

BALB/c

1. Fresh tumor tissues derived from HCC patients were inoculated into the back subcutaneous of 6-8-week-old NSG mice, then generated the PDX model.

2. HepG2 cells were transplanted into the dorsal flanking of 6-8-week-old male BALB/c nude mice. Then, tumor metastases to mouse liver were selected for the experiments.

Hu et al. [97]

implantation mouse models

BALB/c

Fresh HCC tissues from clinical surgical specimens were cut to a depth of 2 mm in diameter and subcutaneously buried in the right axilla of eighty nude mice by a trocar puncture. The mice were continuously fed, and the growth of the tumors was regularly observed. The tumor xenograft model was observed for 35 days. the tumors in the control group exceeded the criteria (3000 mm³) defined by the experimental animal ethics committee, the observation was terminated.

Nazzal et al. [98]

implantation mouse models

NOD/SCID

HCC liver specimens from HCC patient were cut into small pieces (1–3 mm³) and directly implanted into NSG mice. Mice were anesthetized using isoflurane (1–3%), skin aseptically prepared, and a small dorsal midline incision (< 10 mm) was made at the level of the flank. Tumor tissues were placed in bilateral subcutaneous pockets and the incision was closed. Lidocaine was infiltrated at the wound edges to control postoperative pain. When tumor volume reached > 600 mm³, the mouse was humanely sacrificed and the tumors were cryopreserved or explanted for passage in another NSG mouse using the same protocol. Xenograft tumor was developed after 6–7 weeks from mice and was successfully passaged in mice for three generations.