Application protocol

Isolation, expansion, and phenotyping of umbilical cord-derived mesenchymal stem cells

Mesenchymal stem cells or mesenchymal stromal cells (MSCs), are somatic or adult stem cells that can be isolated from various sources, including bone marrow, adipose tissue or umbilical cord. Isolated MSCs are fibroblast-like cells that demonstrate site-specific differentiation. MSCs may regenerate damaged or diseased tissues in vivo and potentially play a role in immunomodulation, which provides the basis for a variety of clinical applications. In this application protocol, we describe the enumeration, isolation, expansion and quality assessment of MSCs derived from umbilical cord.

Materials

The following is a listing of reagents, instruments, and consumables needed for each step of this protocol. These products are for research use only.

For dissociation of umbilical cord tissue

  • Umbilical Cord Dissociation Kit (# 130-105-737)
  • Phosphate-buffered saline, pH 7.2
  • DMEM
  • gentleMACS™ Octo Dissociator with Heaters (# 130-096-427)
  • gentleMACS C Tubes (# 130-093-237)
  • MACS® SmartStrainers (100 µm) (# 130-098-463)
  • (Optional) 70% ethanol
  • (Optional) MACS Tissue Storage Solution (# 130-100-008)
  • (Optional) ART® 1000 REACH™ pipet tips for removal of dissociated material from the closed C Tubes

For MSC expansion

StemMACS™ MSC Expansion Media Kit XF (# 130-104-182)

For MSC phenotypic and functional analysis

  • StemMACS AdipoDiff Media, human (# 130-091-677)
  • StemMACS ChondroDiff Media, human (# 130-091-679)
  • StemMACS OsteoDiff Media, human (# 130-091-678)
  • MSC Phenotyping Kit, human (# 130-095-198)
  • MSC Suppression Inspector, human (# 130-096-207)

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Protocol


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This protocol includes detailed instructions for preparing necessary reagents and materials and then carrying out each step.