# Understanding and Using Variation in Source Materials for MSC Fabrication

> **NIH NIH R01** · CLEVELAND CLINIC LERNER COM-CWRU · 2021 · $559,242

## Abstract

Project Summary
The Broad Aim of this proposal is to advance the field of stem cell biology, cell-based diagnostics, and cell
sourcing for cell therapy and drug development using a new pathway for systematic isolation of specific stem
cell and progenitor cell subtypes from the plurality of niches in musculoskeletal connective tissues that we refer
to as “Performance-Based Selection” (PBS). Our goal is to provide patients, clinicians and researchers with the
knowledge and rigorous and reproducible tools and standards that they need, to identify and obtain the best
possible cells for use in regenerative and joint preservation therapies, drug discovery and basic research.
Mesenchymal Stromal Cells (MSCs) derived from colony founding Connective Tissue Progenitors (CTPs) in
bone and other tissues are widely valuable for cellular therapy. However, batch to batch MSC variation
represents a profound point of ongoing “pain” in the biomedical research community that is limiting the clinical
evaluation and performance of MSC-based products. Clinical therapies demand high levels of repeatability and
reproducibility. Minimizing variation is essential to advancing the emerging field of cellular therapies.
We hypothesize that a large source of this variation is the failure of traditional methods of MSC fabrication based
on “Competitive Expansion” (CE), where the user simply puts all cells from tissue source into culture, where they
compete. No choice is made regarding cells to include or exclude. The user simply accepts an outcome that may
be predetermined by random variation in the type and mix of colony founding cells in the initial culture.
This proposal integrates several unique technology platforms to enable rapid and effective progress in
overcoming these limitations, specifically:
1) Standardized robotics for automated large-field-of-view (LFOV) (“Google Earth”) imaging and image analysis
 to identify and measure CTPs and their progeny.
2) Time Lapse Videomicroscopy that enables continuous tracking of the formation of each CTP-derived colony
 from the time of plating. Reversing the video enables us, for the first time, to see and to characterize colony
 founding CTPs as they exist immediately after removal from our bodies and before they divide.
3) An automated robotic platform (Cell X™) that enables both the selection (“Picking”) of individual CTP-derived
 clones as well as the automated processing that is needed to expand their progeny “hands free”, enabling
 vastly greater precision and reproducibility than current manual methods.
This proposal builds the knowledge that CTPs vary widely in biological potential. Using these tools, we will
determine the attributes of colony founding CTPs that should or should not be included in MSC manufacturing
and then provide the means to use this knowledge to achieve high quality and highly reproducible outcomes.

## Key facts

- **NIH application ID:** 10165695
- **Project number:** 5R01DE029634-03
- **Recipient organization:** CLEVELAND CLINIC LERNER COM-CWRU
- **Principal Investigator:** George F Muschler
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $559,242
- **Award type:** 5
- **Project period:** 2019-08-14 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10165695

## Citation

> US National Institutes of Health, RePORTER application 10165695, Understanding and Using Variation in Source Materials for MSC Fabrication (5R01DE029634-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10165695. Licensed CC0.

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