人乳腺癌细胞 disease models

Use case

Use MDA-MB-231 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 MDA-MB-231 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 (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 6272987Cancer Res · 1981
Characterization of a new human breast carcinoma cell line (MDA-MB-231).
PubMedPMID 6606682J Immunol Methods · 1983
Rapid colorimetric assay for cellular growth and survival.

Use case

Assess MDA-MB-231 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 (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 22130855J Vis Exp · 2011
In vitro cell migration and invasion assays.
PubMedPMID 17546025Cell Adh Migr · 2007
An introduction to the wound healing assay using live-cell microscopy.

Use case

Low-density plating to score colonies from single cells—better than short MTT/CCK-8 for long-term proliferative capacity and post-treatment clonogenic survival.

In one line

Count → low-density seed → ± drug → 10–14 days → fix/stain → score colonies (≥50 cells).

Protocol overview

01
Expand; log passage
02
Trypsinize; accurate count
03
Low-density 6-well seed
04
Attach ± drug / pretreat
05
Feed 10–14 days
06
Methanol + crystal violet
07
Score colonies; survival fraction

Reagents / compounds

Name	Role	Conc.
Test compound	Drug / clonogenic survival	对数梯度或固定暴露窗，预实验优化
Vehicle	Vehicle control	等体积分数
Crystal violet	Colony stain	约 0.5%（按 SOP）

Readouts

Colony counts, plating efficiency (PE), survival fraction (SF), optional colony area.

Literature (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 17487121Nat Protoc · 2006
Clonogenic assay of cells in vitro.
PubMedPMID 33339004Molecules · 2020
Comparative study of lipophilic statin activity in 2D and 3D models of MDA-MB-231 and MCF-7.

Use case

Separate viability loss into apoptosis, necrosis, or cell-cycle arrest. Annexin V–PI, caspase activity, and PI/EdU are common MDA-MB-231 readouts.

In one line

Seed → dose → harvest adherent+floaters → Annexin V / cycle stain → flow; paired viability.

Protocol overview

01
Seed to assay density
02
Dose + vehicle
03
Incubate 24–72 h
04
Harvest adherent + floaters
05
Annexin V–PI or caspase
06
Parallel PI/EdU cycle
07
Gate and quantify by flow

Reagents / compounds

Name	Role	Conc.
Test compound	Apoptosis / cycle perturbation	预实验优化
Annexin V–PI (or 7-AAD) kit	Apoptosis gating	按说明书
PI + RNase (or EdU)	Cell-cycle analysis	按说明书
Staurosporine (optional positive control)	Apoptosis positive control	低 nM–μM，预实验

Readouts

% early/late apoptosis, sub-G1, G1/S/G2 distribution, paired viability.

Literature (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 25374279Oncol Rep · 2015
Andrographolide inhibits MDA-MB-231 migration/invasion and induces apoptosis via NF-κB/MMP-9.
PubMedPMID 6606682J Immunol Methods · 1983
Rapid colorimetric assay for cellular growth and survival.

Use case

Ultra-low-attachment or hanging-drop spheroids to mimic 3D context and drug penetration; MDA-MB-231 2D vs 3D drug responses often diverge.

In one line

Count → ULA/hanging-drop → confirm spheroids → ± drug → diameter/viability → optional section or dissociate.

Protocol overview

01
Accurate single-cell count
02
Seed ULA or hanging drop
03
Optional spin to aggregate
04
Form 2–4 days
05
Image compact spheres
06
Dose; track diameter
07
Endpoint ATP/CTG or counts

Reagents / compounds

Name	Role	Conc.
Test compound	3D drug response	常高于 2D EC₅₀，预实验优化
Vehicle	Control	等体积分数
3D viability reagent (e.g. CTG 3D)	Spheroid ATP/viability	按说明书

Readouts

Spheroid diameter/area, ATP viability, optional necrotic core and dissociated counts.

Literature (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 33339004Molecules · 2020
Comparative study of lipophilic statin activity in 2D and 3D models of MDA-MB-231 and MCF-7.
PubMedPMID 19214182Nat Protoc · 2009
Spheroid-based drug screen: considerations and practical approach.

Use case

Orthotopic mammary fat-pad tumors in immunocompromised mice, or tail-vein lung / intratibial bone metastasis—core MDA-MB-231 in vivo readouts. Follow IACUC and biosafety rules.

In one line

Expand early passage → viable count → choose injection route → monitor tumor/weight/imaging → endpoint lung/bone pathology.

Protocol overview

01
IACUC + animal quarantine
02
Expand MDA-MB-231 (early passage)
03
Count; viability ≥90%
04
Route: orthotopic / i.v. / intratibial
05
Inject + postop care
06
Caliper or bioluminescence
07
Endpoint lung/bone pathology

Reagents / compounds

Name	Role	Conc.
MDA-MB-231 suspension	Inoculum	按路径优化（原位约 10⁶；静脉常 10⁵ 级）
Matrigel (optional orthotopic)	Engraftment aid	按 SOP 混合
Test drug / vehicle	Treatment arm	按药理与伦理批件

Readouts

Tumor volume, body weight, lung colonies, osteolysis/pathology, optional BLI/fluorescence.

Literature (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 6272987Cancer Res · 1981
Characterization of a new human breast carcinoma cell line (MDA-MB-231).
PubMedPMID 1692766Cancer Res · 1990
Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice.
PubMedPMID 25374279Oncol Rep · 2015
Andrographolide inhibits MDA-MB-231 progression and breast cancer bone metastasis model readouts.

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 (PubMed)

PubMedPMID 4357757J Natl Cancer Inst · 1973
Breast tumor cell lines from pleural effusions.
PubMedPMID 6272987Cancer Res · 1981
Characterization of a new human breast carcinoma cell line (MDA-MB-231).

triple-negative breast adenocarcinoma 2D viability / drug screen — foolproof

Use case

In one line

Foolproof steps (8)

Reagents / compounds

Readouts

Literature (PubMed)

Migration / invasion — foolproof

Use case

In one line

Foolproof steps (8)

Reagents / compounds

Readouts

Literature (PubMed)

Clonogenic / colony-formation — foolproof

Use case

In one line

Foolproof steps (9)

Reagents / compounds

Readouts

Literature (PubMed)

Apoptosis / cell-cycle — foolproof

Use case

In one line

Foolproof steps (9)

Reagents / compounds

Readouts

Literature (PubMed)

3D spheroid culture — foolproof

Use case

In one line

Foolproof steps (9)

Reagents / compounds

Readouts

Literature (PubMed)

Orthotopic / lung or bone metastasis (in vivo) — foolproof

Use case

In one line

Foolproof steps (9)

Reagents / compounds

Readouts

Literature (PubMed)

Passage-window & QC checklist

Use case

In one line

Foolproof steps (6)

Readouts

Literature (PubMed)