# scientific experiment protocol

## Pump.Science Longevity Testing Protocol (Simple Overview)

This document explains our three‑stage pipeline for evaluating compounds that may extend lifespan and improve healthspan. Each stage builds on the previous one, starting with short‑lived invertebrates and progressing to mammals.

***

### Stage 1 – **C. elegans Lifespan Assay**

| Item             | Details                                                                                   |
| ---------------- | ----------------------------------------------------------------------------------------- |
| Contract Lab     | **Ora Biomedical**                                                                        |
| Model Organism   | *Caenorhabditis elegans* (wild‑type N2)                                                   |
| Group Size       | 60 worms / compound vs 60 worms / vehicle control                                         |
| Primary Endpoint | **Median & maximum lifespan**                                                             |
| Runtime          | Up to **8 weeks** (culture → final death event)                                           |
| Data Delivery    | Experiment is run first; full video & data are **streamed retrospectively** once complete |

**Why this matters**  C. elegans provide a rapid first screen for pro‑longevity effects before investing in more complex organisms.

***

### Stage 2 – **Drosophila (Fly) Health & Lifespan Assay**

| Item              | Details                                                                                                                                     |
| ----------------- | ------------------------------------------------------------------------------------------------------------------------------------------- |
| Contract Lab      | **Tracked Biotechnologies**                                                                                                                 |
| Model Organism    | *Drosophila melanogaster*                                                                                                                   |
| Group Size        | 45 flies / compound vs 45 flies / control                                                                                                   |
| Primary Endpoints | <p>• Lifespan• <strong>Movement metrics</strong> (speed & total distance)</p><p>• <strong>Negative geotaxis</strong> (climbing latency)</p> |
| Lead Time         | **≈ 10 days** to culture and load arenas                                                                                                    |
| Runtime           | **≈ 70 days** total                                                                                                                         |
| Data Delivery     | Continuous **streaming video** + rolling data uploads                                                                                       |

**Why this matters**  Motor performance (geotaxis) and spontaneous activity act as surrogates for overall health, revealing functional benefits beyond survival alone.

***

### Stage 3 – **Mouse Lifespan & Functional Assay**

| Item                  | Details                                                                                                    |
| --------------------- | ---------------------------------------------------------------------------------------------------------- |
| Contract Lab          | **VivoArchitect**                                                                                          |
| Model Organism        | C57BL/6 mice                                                                                               |
| Group Size            | 20 mice / compound vs 20 mice / control                                                                    |
| Primary Endpoint      | **Overall survival (lifespan)**                                                                            |
| Supplementary Tests\* | Open Field, Rotarod, Y‑Maze, Elevated Plus Maze, Hanging Test, Grip Strength, Automated Gender Recognition |
| Health Monitoring     | Body weight, rectal temperature, CBC, plasma safety biomarkers                                             |
| Sourcing              | Mice purchased from licensed SPF vendor (C57BL/6, 6‑8 weeks old)                                           |

\*Functional assays are selected case‑by‑case, driven by the compound’s proposed mechanism.

***

### Stage 4 – **Human Wearable‑Based Double‑Blind Study**

| Item                | Details                                                                                         |
| ------------------- | ----------------------------------------------------------------------------------------------- |
| Study Type          | Randomised, placebo‑controlled, double‑blind                                                    |
| Participants        | \~30 healthy adults per arm (exact N defined per protocol)                                      |
| Primary Endpoints   | Wearable‑derived endpoints, i.e. **VO₂max**, **resting heart rate**, **sleep efficiency, etc.** |
| Secondary Endpoints | Heart‑rate variability, activity level, other validated wearable biomarkers                     |
| Runtime             | **12 weeks**                                                                                    |
| Data Delivery       | De‑identified wearable data streamed to an open dashboard in near‑real time                     |

**Why this matters**  Human data confirms translatability and identifies early signals of safety & efficacy before larger trials.

***

* **Compound Dosing** – Dosing regimens are set in consultation with each CRO and compound submitter.

For detailed, peer‑reviewable methodology, refer to our [preprint](https://www.researchhub.com/paper/9427260/automated-transparent-and-scalable-longevity-research-across-model-organisms-introducing-the-pumpscience-protocol)

***

*Last updated: 2025‑07‑06*


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