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. 2020 Sep 14;2(5):e629-e636.
doi: 10.1016/j.asmr.2020.07.013. eCollection 2020 Oct.

Analysis of Time to Form Colony Units for Connective Tissue Progenitor Cells (Stem Cells) Harvested From Concentrated Bone Marrow Aspirate and Subacromial Bursa Tissue in Patients Undergoing Rotator Cuff Repair

Arthur Landry  1 , Benjamin J Levy  2 , Mary Beth McCarthy  2 , Lukas N Muench  2   3 , Colin Uyeki  2 , Daniel P Berthold  2   3 , Mark P Cote  2 , Augustus D Mazzocca  2
Affiliations

Analysis of Time to Form Colony Units for Connective Tissue Progenitor Cells (Stem Cells) Harvested From Concentrated Bone Marrow Aspirate and Subacromial Bursa Tissue in Patients Undergoing Rotator Cuff Repair

Arthur Landry et al. Arthrosc Sports Med Rehabil. .

Abstract

Purpose: To evaluate the time required for colonies to develop from concentrated bone marrow aspirate (cBMA) and subacromial bursal tissue samples.

Methods: Samples of cBMA and subacromial bursa tissue were harvested from patients undergoing rotator cuff repair surgery between November 2014 and December 2019. Samples were analyzed for time to form colonies and number of colonies formed. The impact of age, sex, and cellularity (cBMA only) was analyzed. Samples were cultured and evaluated daily for colony formation in accordance with the guidelines of the International Society for Cellular Therapy. Demographic factors were analyzed for impact on time to form colonies and number of colonies formed.

Results: Samples of cBMA were obtained from 92 patients. Subacromial bursa tissue was obtained from 54 patients. For cBMA, older age was associated with more days to form colonies (P = .003), but sex (P = .955) and cellularity (P = .623) were not. For bursa, increased age was associated with longer time to form colonies (P = .002) but not sex (P = .804). Conclusions: Increased age (in cBMA and subacromial bursa tissue) and lower initial cellularity (in cBMA) are associated with longer time to form colonies in culture.

Clinical relevance: Although connective tissue progenitor cells are widely used in orthopaedic practice, there are few metrics to determine their efficacy. Time to form colonies may serve as an important measurement for determining connective tissue progenitor cell viability for augmentation of rotator cuff repair. Subacromial bursa tissue may represent a viable alternative to cBMA for augmentation of rotator cuff repair, capable of forming colonies expediently in vivo.

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Figures

Fig 1
Fig 1
Time-lapse photography (10× magnification) of bursa and cBMA cells plated in culture taken at 24-hour intervals. At 24 hours after plating (A and B), cells in culture before colonies have formed, overlapping cells and lack of colony groups. Colonies are noted to begin forming in images C and D (48 hours after plating), indicated by circled areas. Images E and F (72 hours after plating) show cells approaching confluence, making individual colony counting impossible. (cBMA, concentrated bone marrow aspirate.)
Fig 2
Fig 2
Presence of confirmatory surface markers (CD73, CD90, and CD105) for presence of CTPs in cBMA and subacromial bursa samples. FACS analysis of 8 random cBMA and subacromial bursa samples were performed for the aforementioned analysis. (cBMA, concentrated bone marrow aspirate; FACS, fluorescence-activated cell sorting.)
Fig 3
Fig 3
Days to form colonies versus age for cBMA and bursa. (cBMA, concentrated bone marrow aspirate.)
Fig 4
Fig 4
Total CFUs formed for cBMA and bursa. (cBMA, concentrated bone marrow aspirate; CFU, colony-forming units.)
Fig 5
Fig 5
Average time to form colonies (days) for bursa and cBMA. (cBMA, concentrated bone marrow aspirate.)
Fig 6
Fig 6
Total colonies formed (CFU) for bursa and cBMA. (cBMA, concentrated bone marrow aspirate; CFU, colony-forming units.)
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