人子宫内膜癌细胞 disease models

Ishikawa (Type I endometrial adenocarcinoma). Standardized family protocols with foolproof steps, flowcharts, and methodology PubMed refs—optimize per datasheet and pilots. Research use only (RUO).

Adapted from PubMed literature for research use only (RUO); adjust per lab SOP and pilots.

Use case

Use Ishikawa in 2D culture to test how compounds or genetic perturbations affect proliferation/survival—baseline solid-tumor readout.

In one line

Stable culture → seed overnight → dose matrix → timed viability/apoptosis → compare vs vehicle.

Protocol overview

01
Expand; log passage
02
Seed plates to assay density
03
Attach / stabilize overnight
04
Add dose matrix + vehicle
05
Incubate to endpoint
06
Viability / apoptosis assay
07
Compute IC₅₀ or % inhibition

01[Prep] Confirm complete medium per datasheet; mycoplasma-negative before key assays.
02[Seed] Count and plate so cultures are ~40–70% confluent at dosing; overnight for adherent lines.
03[Dose] Vehicle + ≥5-point log dose grid; optional cytotoxic positive control. Keep DMSO ≤0.1–0.5% if possible.
04[Incubate] Typically 24–72 h; minimize disturbance.
05[Viability] MTT / CCK-8 / CellTiter-Glo per kit; ≥3 wells/group.
06[Optional apoptosis] Annexin V / caspase or DNA fragmentation.
07[Optional colony] Low-density plating, 1–3 weeks, crystal violet colony counts.
08[Calculate] % inhibition vs vehicle; fit IC₅₀; ≥2–3 independent repeats.
09[QC] Log passage and serum lot; exclude contaminated/detached wells.

Reagents / compounds

Name	Role	Conc.
Test compound / inhibitor	Drug response or pathway intervention	对数梯度；按文献与预实验优化
Vehicle control (e.g. DMSO)	Rule out vehicle artifacts	与最高药组等体积分数
Viability reagent (MTT / CCK-8 / CTG)	Quantify viability/metabolic activity	按试剂盒说明书

Readouts

(1) Viability / IC₅₀; (2) optional apoptosis; (3) optional colonies; (4) morphology.

Literature & manuals

[1]PubMedPMID 4031568Nihon Sanka Fujinka Gakkai zasshi · 1985
[Establishment of a new human endometrial adenocarcinoma cell line, Ishikawa cells, containing estrogen and progesterone receptors].
[2]PubMedPMID 10381137Cancer letters · 1998
In vitro effects of gonadotropin-releasing hormone (GnRH) analogue on cancer cell sensitivity to cis-platinum.
[3]PubMedPMID 6606682J Immunol Methods · 1983
Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.
[4]PubMedPMID 16740937Nat Protoc · 2006
Clonogenic assay of cells in vitro.
[5]PubMedPMID 2135685J Exp Med · 1990
Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis.

Use case

Assess horizontal migration or matrix invasion of Ishikawa for metastasis-related mechanisms and anti-migratory screens.

In one line

Confluent scratch or Transwell seeding → treat → timed imaging / count invaded cells.

Protocol overview

01
Seed to confluence or Transwell
02
Scratch or add invasion matrix
03
Switch to drug + low-serum medium
04
Timed imaging / stain membrane
05
Quantify wound area or invaded cells
06
Compare vs vehicle

Reagents / compounds

Name	Role	Conc.
Low-serum medium	Reduce proliferation confounders	常用 0.5–2% FBS（按 SOP）
Matrigel / invasion matrix (optional)	Model ECM invasion	按说明书稀释铺胶
Test compound / inhibitor	Drug response or pathway intervention	对数梯度；按文献与预实验优化
Vehicle control (e.g. DMSO)	Rule out vehicle artifacts	与最高药组等体积分数

Readouts

(1) Wound closure; (2) invaded cell counts; (3) paired viability.

Literature & manuals

[1]PubMedPMID 4031568Nihon Sanka Fujinka Gakkai zasshi · 1985
[Establishment of a new human endometrial adenocarcinoma cell line, Ishikawa cells, containing estrogen and progesterone receptors].
[2]PubMedPMID 10381137Cancer letters · 1998
In vitro effects of gonadotropin-releasing hormone (GnRH) analogue on cancer cell sensitivity to cis-platinum.
[3]PubMedPMID 17546025Cell Adh Migr · 2007
An introduction to the wound healing assay using live-cell microscopy.
[4]PubMedPMID 22130855J Vis Exp · 2011
In vitro cell migration and invasion assays.

Use case

Fix passage window and mycoplasma/morphology baselines to reduce lot drift.

In one line

Log thaw passage → mycoplasma check → archive morphology → run key assays in-window.

Protocol overview

01
Thaw & log passage
02
Mycoplasma check
03
Archive morphology
04
Run key assays in window

Readouts

Mycoplasma-negative, stable morphology, in-window passage, viability baseline.

Literature & manuals

[1]PubMedPMID 4031568Nihon Sanka Fujinka Gakkai zasshi · 1985
[Establishment of a new human endometrial adenocarcinoma cell line, Ishikawa cells, containing estrogen and progesterone receptors].
[2]PubMedPMID 10381137Cancer letters · 1998
In vitro effects of gonadotropin-releasing hormone (GnRH) analogue on cancer cell sensitivity to cis-platinum.

Type I endometrial adenocarcinoma-related 2D viability / drug screen — foolproof protocol

Use case

In one line

Foolproof steps (9)

Reagents / compounds

Readouts

Literature & manuals

Migration / invasion — foolproof protocol (scratch or Transwell)

Use case

In one line

Foolproof steps (8)

Reagents / compounds

Readouts

Literature & manuals

Passage-window & QC checklist

Use case

In one line

Foolproof steps (6)

Readouts

Literature & manuals