{"title":"Xenogen-free media provide variable equine mesenchymal stromal cell expansion after a 7-day culture period.","authors":"Maureen K Larson, Caitlin Gaffney, Cherise Hoagland, Judith Jayawickrama, J Lacy Kamm","doi":"10.2460/ajvr.25.03.0109","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To determine the xenogen-free serum source that provides the greatest number of live equine mesenchymal stromal cells (MSCs) while maintaining the MSC phenotype.</p><p><strong>Methods: </strong>Equine bone marrow-derived MSCs from 8 horses were cultured for 7 days in media containing one of the following serum treatments: 10% xenogeneic serum, 10% or 20% commercial allogeneic equine serum, 10% autologous serum, 10% equine pooled platelet lysate (PPL), or a staged media reduction of xenogeneic media. Live cell numbers, MSC viability, and MSC immunophenotype were compared.</p><p><strong>Results: </strong>The use of 10% commercial allogeneic equine serum in media resulted in a significant decrease in live MSC count compared to xenogeneic serum (P = .01; 95% CI, -244,766 to -48,038). Autologous serum, staged reduction, and PPL had no significant difference in live MSC number collected at day 7 as compared to xenogeneic serum. There were no significant differences in cell count due to the horse's age group. Viability was significantly greater using PPL than all other xenogen-free serums across the < 10-year and ≥ 10-year age groups. Mesenchymal stromal cells from all treatments were high in CD44, CD90, and major histocompatibility complex (MHC) I expression and low in CD45 and MHC II expression. Significant differences in CD44, CD45, CD90, MHC I, and MHC II expression levels were seen across treatment and age analysis.</p><p><strong>Conclusions: </strong>The use of PPL for MSC media in the final 7 days of culture allows for adequate expansion and viability of MSCs and maintenance of MSC immunophenotypic markers.</p><p><strong>Clinical relevance: </strong>MSCs can be cultured in nonxenogeneic media for 7 days prior to harvest.</p>","PeriodicalId":7754,"journal":{"name":"American journal of veterinary research","volume":" ","pages":"1-9"},"PeriodicalIF":1.4000,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"American journal of veterinary research","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.2460/ajvr.25.03.0109","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"VETERINARY SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: To determine the xenogen-free serum source that provides the greatest number of live equine mesenchymal stromal cells (MSCs) while maintaining the MSC phenotype.
Methods: Equine bone marrow-derived MSCs from 8 horses were cultured for 7 days in media containing one of the following serum treatments: 10% xenogeneic serum, 10% or 20% commercial allogeneic equine serum, 10% autologous serum, 10% equine pooled platelet lysate (PPL), or a staged media reduction of xenogeneic media. Live cell numbers, MSC viability, and MSC immunophenotype were compared.
Results: The use of 10% commercial allogeneic equine serum in media resulted in a significant decrease in live MSC count compared to xenogeneic serum (P = .01; 95% CI, -244,766 to -48,038). Autologous serum, staged reduction, and PPL had no significant difference in live MSC number collected at day 7 as compared to xenogeneic serum. There were no significant differences in cell count due to the horse's age group. Viability was significantly greater using PPL than all other xenogen-free serums across the < 10-year and ≥ 10-year age groups. Mesenchymal stromal cells from all treatments were high in CD44, CD90, and major histocompatibility complex (MHC) I expression and low in CD45 and MHC II expression. Significant differences in CD44, CD45, CD90, MHC I, and MHC II expression levels were seen across treatment and age analysis.
Conclusions: The use of PPL for MSC media in the final 7 days of culture allows for adequate expansion and viability of MSCs and maintenance of MSC immunophenotypic markers.
Clinical relevance: MSCs can be cultured in nonxenogeneic media for 7 days prior to harvest.
期刊介绍：
The American Journal of Veterinary Research supports the collaborative exchange of information between researchers and clinicians by publishing novel research findings that bridge the gulf between basic research and clinical practice or that help to translate laboratory research and preclinical studies to the development of clinical trials and clinical practice. The journal welcomes submission of high-quality original studies and review articles in a wide range of scientific fields, including anatomy, anesthesiology, animal welfare, behavior, epidemiology, genetics, heredity, infectious disease, molecular biology, oncology, pharmacology, pathogenic mechanisms, physiology, surgery, theriogenology, toxicology, and vaccinology. Species of interest include production animals, companion animals, equids, exotic animals, birds, reptiles, and wild and marine animals. Reports of laboratory animal studies and studies involving the use of animals as experimental models of human diseases are considered only when the study results are of demonstrable benefit to the species used in the research or to another species of veterinary interest. Other fields of interest or animals species are not necessarily excluded from consideration, but such reports must focus on novel research findings. Submitted papers must make an original and substantial contribution to the veterinary medicine knowledge base; preliminary studies are not appropriate.
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