Sequential sub-passage decreases the differentiation potential of canine adipose-derived mesenchymal stem cells.
Adipose-derived mesenchymal stem cells (AD-MSCs) are abundant in adipose tissue from animals of all ages, are easily isolated, can differentiate into multi-lineage cells, and have a clinical application. This promising potential may only be achieved if the cells are expanding in a large number while maintaining their stemness in sequential passages. In this study, canine AD-MSCs (cAD-MSCs) were individually isolated from five dogs and subjected to proliferative culture with seven sub-passages. The cells at each sub-passage were characterized for properties associated with multipotent MSCs such as proliferation kinetics, expression of MSCs-specific surface markers, expression of molecules associated with self-renewal and differentiation capabilities into mesodermal lineage cells. Proliferation of the cells plateaued at passage 5 by cumulative population doubling level, while cell doubling time gradually increased with passage. MSCs surface markers (CD44, CD90, and CD105) and molecules (Oct 3/4, Sox-2, Nanog and HMGA2) associated with self-renewal were all expressed in the cells between passages 1 to 6 by RT-PCR. In addition, the cells at passage 1, 3 or 6 underwent adipogenic and chondrogenic differentiation under specific induction conditions. However, the level of adipogenic and chondrogenic differentiation was negatively correlated with the number of sub-passage. The present study suggests that sequential sub-passages affect multipotent properties of cAD-MSCs, which should be considered in their therapeutic application in regenerative medicine.
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea; Department of Animal Biotechnology, KonKuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
Keum Sil
KS
Lee
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea.
Hye Won
HW
Kang
Department of Animal Biotechnology, KonKuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
Hoon Taek
HT
Lee
Haemaru Referral Animal Hospital, Bundang-gu, Seongnam-si, Gyeonggi-do 463-824, Republic of Korea.
Hye-Jin
HJ
Kim
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea.
Chan-Lan
CL
Kim
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea.
Jae-Young
JY
Song
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea. Electronic address: kwlee@korea.kr.
Kyung Woo
KW
Lee
Animal and Plant Quarantine Agency, Anyang 6-dong, Anyang-si, Gyeonggi-do 430-757, Republic of Korea. Electronic address: virusmania@korea.kr.
Sang-Ho
SH
Cha
D050156:Adipogenesis / Q000276:immunology; D000273:Adipose Tissue / Q000166:cytology* / Q000276:immunology; D000818:Animals; D002454:Cell Differentiation / Q000276:immunology*; D020219:Chondrogenesis / Q000276:immunology; D004285:Dogs / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D025743:HMGA2 Protein / Q000235:genetics / Q000276:immunology; D018398:Homeodomain Proteins / Q000235:genetics / Q000276:immunology; D016130:Immunophenotyping / Q000379:methods / Q000662:veterinary; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D050813:Octamer Transcription Factor-2 / Q000235:genetics / Q000276:immunology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000276:immunology
D025743:HMGA2 Protein; D018398:Homeodomain Proteins; D050813:Octamer Transcription Factor-2; D055748:SOXB1 Transcription Factors
Adipose tissue; Canine; Differentiation; Mesenchymal stem cell; Multipotent