人结肠癌细胞 disease models

HT-29 (colorectal adenocarcinoma) classic parental modeling page with foolproof steps, flowcharts, and PubMed refs. Can differentiate toward enterocyte-like phenotype under defined conditions. Optimize per datasheet/pilots; RUO.

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

Use case

Use HT-29 to test compound/genetic effects on proliferation/survival.

In one line

Expand → seed → dose matrix → viability/apoptosis.

Protocol overview

01
Expand; log passage
02
Seed overnight
03
Dose + vehicle
04
Incubate 24–72 h
05
Viability assay
06
Fit IC₅₀
07
Independent repeats

01[Prep] Confirm HT-29 medium; mycoplasma-negative.
02[Seed] Plate to ~40–70% confluence at dosing; overnight attach.
03[Dose] Vehicle + ≥5-point log grid; DMSO ≤0.1–0.5% if possible.
04[Incubate] Typically 24–72 h.
05[Viability] MTT/CCK-8/CTG; ≥3 wells/group.
06[Optional] Apoptosis or colony formation.
07[Calculate] % inhibition / IC₅₀; ≥2–3 repeats.
08[QC] Log passage and serum lot.

Reagents / compounds

Name	Role	Conc.
Test compound	Drug/mechanism	对数梯度，预实验优化
Vehicle	Vehicle control	等体积分数
Viability reagent	Quantify viability	按说明书

Readouts

Viability, IC₅₀, optional apoptosis/colonies.

Literature & manuals

[1]PubMedPMID 4357756Cancer Res · 1973
Establishment and characterization of human colonic adenocarcinoma cell lines.
[2]PubMedPMID 7226224Biol Cell · 1980
Enterocytic differentiation of a human colorectal adenocarcinoma cell line HT-29.
[3]PubMedPMID 6606682J Immunol Methods · 1983
Rapid colorimetric assay for cellular growth and survival.

Use case

Assess HT-29 migration/invasion for metastasis-related work.

In one line

Scratch or Transwell → treat → image/count.

Protocol overview

01
Confluence or Transwell
02
Scratch/matrix
03
Low-serum ± drug
04
Timed imaging
05
Quantify
06
Paired viability

Reagents / compounds

Name	Role	Conc.
Low-serum medium	Migration window	0.5–2% FBS（优化）
Matrigel (optional)	Invasion matrix	按说明书

Readouts

Wound closure, invaded counts, viability.

Literature & manuals

[1]PubMedPMID 4357756Cancer Res · 1973
Establishment and characterization of human colonic adenocarcinoma cell lines.
[2]PubMedPMID 22130855J Vis Exp · 2011
In vitro cell migration and invasion assays.
[3]PubMedPMID 17546025Cell Adh Migr · 2007
An introduction to the wound healing assay using live-cell microscopy.

Use case

Fix passage window and mycoplasma/morphology baselines.

In one line

Log thaw passage → mycoplasma → morphology → in-window assays.

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.

Literature & manuals

[1]PubMedPMID 4357756Cancer Res · 1973
Establishment and characterization of human colonic adenocarcinoma cell lines.
[2]PubMedPMID 7226224Biol Cell · 1980
Enterocytic differentiation of a human colorectal adenocarcinoma cell line HT-29.

colorectal adenocarcinoma 2D viability / drug screen — foolproof

Use case

In one line

Foolproof steps (8)

Reagents / compounds

Readouts

Literature & manuals

Migration / invasion — foolproof

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